Microbiology and Molecular Biology Reviews, June 2000, p. 239-280, Vol. 64, No. 2
University of Texas Southwestern Medical
Center, Dallas, Texas,1 and
University of Washington, Seattle, Washington2
1092-2172/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Translational Control of Viral Gene Expression
in Eukaryotes
SUMMARY
INTRODUCTION
Overview of Eukaryotic mRNA Translation and Sites of
Viral Regulation
Viral translational programming.
Translation initiation.
Cap-binding reaction.
Ribosome scanning and AUG site selection.
Elongation.
Termination.
Improving translation efficiency: the closed-loop model of
mRNA translation.
TRANSLATIONAL CONTROL OF VIRAL GENE EXPRESSION
Advantages and liabilities of cap-dependent host
translation.
Host Shutoff and Selective Translation of Viral mRNA
Mechanisms and Control of Viral mRNA Translation
Internal ribosome entry.
Ribosome shunt.
Leaky scanning.
Frameshifting.
Control of termination and reinitiation.
Functional recoding.
Coupling the Virus Life Cycle to Translational Control
The herpesviruses.
HSV and the shutoff of host cell protein synthesis.
Selective repression of mRNA translation initiation
during HSV infection.
Selective translation of HSV mRNA.
Inhibition of eIF2
phosphorylation
during HSV infection.
Implications of viral modulation of translation in HSV
pathogenesis and disease treatment.
Recruitment of Host Factors for the Efficient
Translation of Viral mRNA
IRES binding proteins and proteins that bind the viral 3'
UTR.
Influenza virus.
Selective translation of influenza virus mRNAs.
Contribution of influenza virus mRNA structure upon
selective translation.
Influenza virus recruitment of Grsf-1.
Temporal regulation of influenza virus mRNA
translation.
Maintenance of translation in influenza virus-infected
cells.
Recruitment of P58IPK and inhibition of eIF2
phosphorylation during influenza virus infection.
Role of the influenza virus NS1 protein in viral
mRNA translation.
VIRAL MODIFICATION OF CELLULAR FACTORS
Inactivation of eIF4E and modulation of the eIF4E-binding
proteins.
Cleavage of eIF4G.
Cleavage of PABP: disruption of the closed-loop translation
complex.
Modification of EF-1.
Disruption of eIF2 phosphorylation.
(i) PKR structure and function.
(ii) Mechanisms of PKR inhibition by eukaryotic viruses.
Disruption of the IFN-induced cellular antiviral response
through inhibition of PKR.
(i) Viral inhibition of PKR: HCV.
VIRAL PERSISTENCE AND TRANSLATIONAL CONTROL
Translational programming and maintenance of viral
persistence.
Translational control, persistent infection, and regulation
of host apoptosis.
Cell growth control, eIF2
phosphorylation, and oncogenic transformation.
MRNA TRANSLATION AS A TARGET FOR
ANTIVIRAL THERAPY
CONCLUSIONS AND PERSPECTIVES
ACKNOWLEDGMENTS
REFERENCES
SUMMARY
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As obligate intracellular parasites, viruses rely exclusively on the translational machinery of the host cell for the synthesis of viral proteins. This relationship has imposed numerous challenges on both the infecting virus and the host cell. Importantly, viruses must compete with the endogenous transcripts of the host cell for the translation of viral mRNA. Eukaryotic viruses have thus evolved diverse mechanisms to ensure translational efficiency of viral mRNA above and beyond that of cellular mRNA. Mechanisms that facilitate the efficient and selective translation of viral mRNA may be inherent in the structure of the viral nucleic acid itself and can involve the recruitment and/or modification of specific host factors. These processes serve to redirect the translation apparatus to favor viral transcripts, and they often come at the expense of the host cell. Accordingly, eukaryotic cells have developed antiviral countermeasures to target the translational machinery and disrupt protein synthesis during the course of virus infection. Not to be outdone, many viruses have answered these countermeasures with their own mechanisms to disrupt cellular antiviral pathways, thereby ensuring the uncompromised translation of virion proteins. Here we review the varied and complex translational programs employed by eukaryotic viruses. We discuss how these translational strategies have been incorporated into the virus life cycle and examine how such programming contributes to the pathogenesis of the host cell.
INTRODUCTION
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Perhaps nowhere in nature is a parasitic relationship as well
defined as that which occurs between a virus and its host cell. Viruses
rely on the host cell for propagation, utilizing cellular machinery for
the replication and assembly of viral components and the release of
progeny virions. Whether possessing a DNA or RNA genome, the eukaryotic
virus exhibits a general life cycle that is initiated through
interaction with its cognate receptor(s) on the surface of the host
cell (117) (Fig. 1). After
virion adsorption and internalization, uncoating exposes the viral
genome and associated proteins to the host milieu, whereupon genome
replication and transcription take place. The translation of viral RNA
is followed by the assembly of structural proteins, packaging of the
viral genome, and eventual release of progeny virions. While some
viruses encode or carry the enzymatic machinery required for autonomous
genome replication and/or transcription, others recruit host
polymerases to carryout this task (117). In contrast, viruses do not encode or carry the machinery for mRNA translation. Thus, the ensuing stage of viral protein synthesis is completely dependent on the translational machinery of the host cell (Fig. 1). Not
surprisingly, viruses have devoted much attention to this dependency
and have evolved strategies that reduce the impact of translational
dependence on viral replication. As discussed in this review, these
strategies are themselves limited by the nature of the viral RNA,
the cellular translation machinery, and the translation regulatory
pathways of the host cell.
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This treatise presents an overview of translation strategies used by viruses that infect the cells of higher eukaryotes. Where appropriate, we have focused on specific virus systems to present examples of the diverse mechanisms by which viruses overcome the problems of translational dependence. For complementary material on mRNA translation, virus-host interactions, host antiviral pathways, and the virus-host interactions of lower eukaryotes and bacteria, we direct the reader to several fine texts and reviews (1, 10, 117, 137, 179, 199, 303, 324, 353, 415, 416, 428, 455, 480, 484). We begin this review with a brief overview of the current models for eukaryotic mRNA translation, including points of translational control, and effects on host translation due to virus infection. This is followed by examples of translation strategies that are dependent on the structure of the viral mRNA and those that are directed at recruitment and modification of the translation machinery and other host factors. Given the recent emphasis in the translational control field on identifying and characterizing cellular signaling pathways that govern mRNA translation (43, 120, 433), we have included discussion of how viruses might exploit these pathways to facilitate completion of their translational programs. Attention is directed to the ways in which disruption of host translational control pathways may contribute to viral pathogenesis and disease progression. Finally, we conclude with a section describing the prospects of targeting viral mRNA translation for antiviral therapy, as well as perspectives for future research in the increasingly overlapping disciplines of virology, viral pathogenesis, and translation control.
Overview of Eukaryotic mRNA Translation and Sites of Viral Regulation
Translation in eukaryotes is a complex multistep, multiprotein process (198, 335). As with most complex biochemical reactions, it is subject to strict regulatory controls, and is extremely sensitive to both the intracellular and extracellular environments (43, 163, 232, 281, 324, 433). In general, the translation of a given mRNA can be modulated in response to nutrient availability, mitogenic stimulation and cell cycle regulation, stress, and, as described herein, viral infection (reviewed in detail in reference 199). It is also increasingly clear from research spanning the past several years that regulation of mRNA translation is critical for maintaining control of cell growth (96, 287, 429). As presented in the "Viral persistence and translational control" section (below), disruption of the major translation checkpoints and signaling cascades renders cells unable to respond to translation-modulatory signals and may constitute a mechanism of oncogenic transformation (67). The following sections provide a general overview of viral translational programming and eukaryotic mRNA translation. Major sites for virus regulation of translation are noted, and they are discussed in detail in this review.
Viral translational programming. Viruses face enormous pressures to maintain a "functional" genome size, which greatly influences the rate and efficiency of viral replication. Thus, host translational dependence may in part reflect the limitations placed on viral replication due to the enormous genome capacity that would be needed to encode the components for autonomous viral protein synthesis (335). This idea is supported by the highly specialized nature of the protein synthetic machinery, which encompasses well over 30 different gene products and yet remains highly conserved between the prokaryotic and eukaryotic kingdoms (335, 470). Eukaryotic viruses have evolved effective means of exploiting their innate translational dependence through mechanisms of translational programming. This is the process in which eukaryotic viruses (i) redirect the host translation machinery to favor viral protein synthesis and (ii) control the expression of their own gene products. The latter is especially important for the RNA viruses, which have limited transcriptional control and rely heavily on translational control strategies to modulate viral gene expression.
Translational programming, such as the use of regulatory upstream open reading frame(s) (uORFs), overlapping reading frames, multicistronic transcripts, and termination control, allows viruses to conserve the functional genome size by making efficient use of genome coding capacity. In general, the mechanisms of translational programming are intrinsic to the structure of the viral mRNA itself. As summarized in Table 1, structural elements within a viral mRNA that affect translational efficiency or impart translational control include the length and structural complexity of the 5' and 3' untranslated regions (UTR), the position and context of the initiator AUG codon, the stability and accessibility of the of the m7G cap and the cap-binding complex, and the presence of uORF(s) preceding the major cistron (137, 148, 149, 269, 270, 322, 329, 430, 431). In addition, cis-acting sequence elements that recruit or bind trans-acting factors can impart an additional level of translational control to viral mRNA by facilitating translational selectivity (23, 76, 187, 244, 361, 362, 394, 411). As described in the following sections, virus translational programming affects all levels of the translation process, including translation initiation, elongation, termination, and host translational control signaling pathways.
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Translation initiation.
The majority of control over
cellular mRNA translation occurs during initiation. Translation
initiation is the process in which the mRNA assembles into a
macromolecular complex with the components required for protein
synthesis, including the eukaryotic initiation factors (eIF) and
elongation factors (EF). Figure 2 shows
the major steps in the cap-dependent translation initiation process and
important sites of virus regulation (for comprehensive reviews of
translation initiation, the reader is referred to references 163, 221, 335, and 359).
Initiation begins with the binding of initiator methionyl-tRNA
(Met-tRNAi) to the 40S ribosomal subunit. This step is
facilitated through the formation of an
eIF2-GTP-Met-tRNAi ternary complex (462). The
recent discovery and functional analyses of eukaryotic homologues of
prokaryotic initiation factor 2 (IF2) indicates that
Met-tRNAi delivery also proceeds via a more general, universally conserved mechanism (60, 291). In this case, IF2 does not participate in the formation of a ternary complex but, rather,
may bind directly to the ribosomal A site and facilitate binding of the
Met-tRNAi to the ribosomal P site during translation initiation. IF2 activity is not subject to direct regulation and therefore may not contribute to the control of mRNA translation. In
contrast, formation of the eIF2-dependent ternary complex and its
delivery of Met-tRNAi to the 40S ribosomal subunit can
constitute a rate-limiting step when the alpha subunit of eIF2
(eIF2
) is phosphorylated by specific protein kinases (see below)
(65, 335). Phosphorylation of eIF2 thus represents a
major point of control over the translation initiation process. eIF2
phosphorylation dramatically alters the efficiency and
rate of mRNA translation and is a critical component of antiviral
and cell growth control pathways (243, 321, 322, 335). eIF2
directs the ternary complex to the 40S ribosomal subunit to form a 43S
pre-initiation complex that includes eIF3 (Fig. 2). eIF3 facilitates
binding of the 43S pre-initiation complex to the mRNA via the
cap-binding complex, eIF4F, that has been assembled around the mRNA
m7G cap structure (335, 359).
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Cap-binding reaction. Assembly of the eIF4F complex on the mRNA is dependent on the eIF4E component of this complex, which recognizes and binds the m7G cap (173). The affinity of eIF4E for the m7G cap constitutes a second major control point in the translation initiation pathway and is subject to variation through eIF4E phosphorylation (237, 315, 433). In addition, the cap-binding activity of eIF4E can be blocked through the formation of an eIF4E-eIF4E binding protein (4E-BP) complex, resulting in inhibition of cap-dependent translation (367, 433). Formation of the eIF4E/4E-BP complex itself is subject to regulation through 4E-BP phosphorylation and dramatically affects cell growth control by altering the efficiency and selectivity of mRNA translation (reviewed by Sonenberg and Gingras [433]). As discussed in detail below, these regulatory steps are targeted by a group of viruses, which are best defined by the family of picornaviruses and includes poliovirus and encephalomyocarditis virus (EMCV) (155, 174, 322). These viruses initiate translation through a cap-independent mechanism that involves internal ribosome entry through use of the internal ribosome entry site (IRES); virus-mediated cleavage of the 220-kDa cap-binding protein, eIF4G (161, 174, 283); and dephosphorylation of the 4E-BPs to sequester eIF4E in an inactive eIF4E/4E-BP complex (Fig. 2) (155). IRES-mediated translation requires specific cis-acting sequences within the viral RNA that mediate the interaction with trans-acting host factors. Thus, the global process of IRES-mediated translation essentially eliminates the competition for host factors from cap-dependent cellular mRNA translation, favoring the translation of viral mRNA.
Ribosome scanning and AUG site selection. Following association with the mRNA, the 43S preinitiation complex begins scanning from the 5' end of the mRNA or the site of ribosome entry (as in the case of cap-independent translation) and continues scanning until the Met-tRNAi interacts with the initiator AUG codon. Ribosomal scanning is not always compatible with mRNAs that possess a long and/or structured 5' UTR. As described in "Mechanism and control of viral mRNA translation" (below), viral mechanisms to overcome the inefficiency of scanning the 5' UTR and to bypass the host shutoff phenomenon include the use of internal ribosomal entry on the mRNA and the ribosomal shunt (93, 223, 494). These mechanisms allow the preinitiation complex to effectively avoid a large part of the 5' UTR and begin scanning within the region of the initiator AUG codon on the viral mRNA (Fig. 2).
Once the Met-tRNAi associates with the initiator AUG codon, GTP is hydrolyzed from the ternary complex, bound initiation factors are released, and the 60S ribosomal subunit joins the preinitiation complex. The resulting 80S initiation complex then mediates the elongation phase of translation. In this model, initiation begins at the 5'-proximal AUG codon (221, 335). However, the AUG site selection for translation initiation is dependent in part on the context of the AUG codon, where the canonical accAUGg sequence (initiation codon in capitals) exerts the highest preference for initiation (221, 270). Departure from this sequence is associated with leaky scanning, in which the preinitiation complex will recognize a noncanonical or weak AUG only at a low frequency and scans past to initiate translation at a downstream codon more closely matching the canonical initiator AUG (Fig. 2) (221). Leaky-scanning initiation of translation is popular among viruses, and in retroviruses it can provide a mechanism for achieving defined stoichiometric ratios of translation products (412, 413).Elongation. During the elongation phase of translation, the mRNA is associated with multiple 80S ribosomes, or polyribosomes, as amino acid residues are sequentially placed on the carboxyl end of the growing peptide chain. In many virus systems the replicative cycle is demarked by early- and late-stage events that can be distinguished by the differential recruitment of viral mRNA into polyribosomal complexes at specific times after infection. As with herpes simplex virus type 1 (HSV-1), this often coincides with the synthesis of latency factors and determinants of virulence (114, 159, 279). The process of translation elongation itself is subject to viral regulatory control (Fig. 2). Elongation control mechanisms include ribosomal frameshifting (108), functional recoding (151), and virus-directed modification of EF-1 (257); the first of these is prevalent in retroviruses and reveals otherwise cryptic ORFs within the viral mRNA (78, 108, 109, 151).
Termination. The process of translation termination occurs when the translating 80S ribosome encounters an in-frame termination codon within the template mRNA. The termination codon is recognized by a release factor, which mediates the hydrolysis of the peptide chain from the bound tRNA (335, 501). This results in the release of the nascent polypeptide from the 80S ribosome and leads to the eventual dissociation of the ribosomal subunits. Once termination has occurred, the 40S subunit is free to continue scanning the mRNA (Fig. 2). In multicistronic transcripts, termination can be followed by reinitiation at the downstream cistron, subject to ternary-complex availability (221). However, reinitiation is usually very inefficient, and the presence of a uORF can confer limitations to the translational efficiency of the major, downstream ORF. As described in "Frameshifting" (below), this termination-reinitiation translational control mechanism is prevalent among viruses and is used to control the synthesis of specific viral gene products (148).
Improving translation efficiency: the closed-loop model of
mRNA translation.
Since the discovery of 3' polyadenylation in
eukaryotic mRNA, it has become quite clear that the poly(A) tail
imparts stimulation of mRNA translation in eukaryotes (reviewed by
Jacobson [225]). More recent analyses indicated that
the translation-stimulatory function of the poly(A) tail was due, in
part, to the actions of the poly(A)-binding protein (PABP). In
mammalian cells, PABP interacts with elements of the cap-binding
complex assembled on the 5' end of the mRNA, thus rendering a
"closed-loop" translation complex (Fig.
3) (138, 225, 401). PABP
promotes the closed loop by binding to eIF4G and to PABP-interacting
protein 1 (Paip-1) (75). Paip-1 interacts with components of
the mRNA cap-binding complex, including eIF4G and the eIF4A
helicase (306). Analyses of translation initiation in yeast
and plants indicate that the interaction between PABP and eIF4G
stimulates mRNA translation (289, 452, 476). The
proximity of the mRNA ends provided by the closed-loop translation
complex is thought to contribute to the stability of the mRNA and
the 5' cap complex and to provide for the efficient recruitment and
recycling of ribosomal subunits (225). Thus, the overall
effect of the closed loop is to increase translation efficiency.
Viruses exploit the closed-loop translation complex as a means of
redirecting the host translation machinery to favor viral mRNA
translation. As described below, viruses accomplish this by targeting
PABP and disrupting the interaction of the mRNA ends, resulting in
attenuation of host mRNA translation (233, 259, 376).
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TRANSLATIONAL CONTROL OF VIRAL GENE EXPRESSION
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Translational dependence has driven viruses to adopt translational programming that maximizes efficiency and facilitates the selective translation of viral mRNA over the endogenous host transcripts. Viral translation strategies have evolved to utilize both the advantages and the limitations inherent within the cap-dependent host translation process. These range from cap-dependent translation competition strategies to cap-independent strategies of IRES-mediated translation initiation. As discussed below, such strategies allow viral mRNA translation to persist, even under the extreme conditions imposed by the host shutoff phenomenon, which severely limits cellular metabolism. This section describes the various translation strategies utilized by eukaryotic viruses to overcome the problems associated with translational dependence and concludes with a discussion of how viral translational programming may present novel targets for the development of anti-viral therapeutics.
Advantages and liabilities of cap-dependent host translation. The majority of mRNA translation within eukaryotic cells is dependent on the m7G cap, a unique structure present at the 5' terminus of the mRNA (335). The 5' cap promotes mRNA stability and nuclear export and provides for various levels of control over the translation initiation process. Cap-dependent control of mRNA translation confers several advantages to the cell. First, and perhaps most importantly, cap dependency allows the cell an immediate mechanism through which to control gene expression by modulating the assembly and activity of cap-binding complex components. Second, cap-dependency provides selectivity of translation by combining the translational regulatory properties inherent within a specific mRNA with those due to modification of the cap-binding complex. Translational control thereby allows the cell to fine-tune gene expression by stimulating or repressing the translation of specific mRNAs, usually through the reversible phosphorylation of translation factors (335).
While cap-dependent translation clearly affords several advantages to the host cell, it also presents liabilities that are effectively exploited by viruses. Cap dependency requires an intact pool of specific initiation factors, namely, the components of the eIF4F cap-binding complex (100, 292, 335, 457, 468). Moreover, it necessitates the capping and nuclear export of mRNAs. Viruses have learned to disrupt these processes in order to reprogram the host cell toward the synthesis of viral proteins. Viral disruption of cap-dependent host translation contributes to the host shutoff that is often observed during productive infections (10).Host Shutoff and Selective Translation of Viral mRNA
Host shutoff is the process in which cellular macromolecular
synthesis is suppressed due to viral domination of host metabolism that
occurs during infection (reviewed in reference 10). Host shutoff is not
an absolute; not all virus infections exhibit host shutoff, and shutoff
is not always required to facilitate viral replication. Within the many
viral systems in which host shutoff is known to occur, shutoff
ultimately favors the translation of viral mRNA over endogenous
host transcripts, although host shutoff itself may not be directly
attributed to viral disruption of host mRNA translation (1,
10). The selective translation of viral mRNA during the host
shutoff is clearly a multicomponent process that has been attributed to
a variety of factors. These include viral perturbation of intracellular
ion concentration (144) and nucleotide metabolism (215,
244, 279), alterations in RNA stability, processing, and export
(119, 245, 311, 352, 497), and the recruitment of specific
host factors (249, 294). From a simpler perspective, the
selective translation of viral mRNA during host shutoff may reflect
a general competition between viral and host mRNA for the
translational machinery. For example, host shutoff in cells infected
with vesicular stomatitis virus (VSV) is coupled to the selective
translation of viral mRNA (Fig. 4, lanes 3 and 4). Interestingly, however, the abundance
and stability of cellular mRNAs and their efficiency of translation
initiation remain unaltered (54). Examination of
VSV-infected cells revealed that the preferential translation of viral
mRNAs was a result of ribosome competition from an overwhelming
abundance of viral mRNA (309). At the other end of the
spectrum is the host shutoff that occurs during
picornavirus infection. In this case, the shutoff of
host protein synthesis and selectivity for viral mRNA translation
is clearly a virus-directed event mediated, in part, through cleavage
of eIF4G by the virus-encoded 2A protease (2A-pro) (174, 273,
468). Cleavage of eIF4G by 2A-pro disrupts cap-dependent
translation initiation to favor the IRES-mediated translation of the
picornavirus mRNA (Fig. 4; also see Fig. 14)
(23). Similarly, the host shutoff in cells infected with
influenza virus features a strong selection for viral mRNA
translation (Fig. 4, lanes 5 and 6). However, unlike the
picornaviruses, influenza virus mRNA translation is
cap dependent (251). In this case, the predominance of viral
protein synthesis is facilitated, in part, by virus-mediated
endonucleolytic cleavage of the host mRNA m7G cap,
subsequent mRNA destabilization, and the
dephosphorylation of eIF4E (244). The
selectivity of viral mRNA translation is then mediated through the
recruitment of the cellular G-rich sequence factor 1 (GRSF-1) protein
and other host factors to the 5' leader sequence of the influenza virus
mRNAs (361, 362).
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Host shutoff can be seen as beneficial for viruses, since it places
cellular resources largely at their disposal. However, the shutoff
phenomenon also presents several challenges to the virus, not least of
which is maintaining the integrity of the host cell long enough to
complete the virus replicative cycle. This is especially important from
the standpoints of translational dependence and viral persistence, in
which the virus must ensure that the host translation machinery remains
competent for the synthesis of viral proteins. Problematically,
however, the metabolic repression and stress of host shutoff are potent
inducers of cellular apoptosis and translational control
programs that function within the cellular antiviral response to block
viral infection (179). Viruses have taken a two-pronged
approach to these problems of host shutoff, and they encode mechanisms
to (i) disrupt host apoptotic programs (353) and (ii)
control the antiviral translational response imposed through the
phosphorylation of eIF2 (65, 66). As described in detail in "Viral modification of cellular factors" (below), eIF2 phosphorylation by the cellular
serine/threonine protein kinase (PKR) presents a translational blockade
to viral replication (68, 131). Disruption of host
apoptosis and the phosphorylation of eIF2
therefore facilitates viral replication by maintaining host cell
integrity and ensuring translational competence during host shutoff.
The relationship between translational control, apoptosis, and
viral infection has been an intense area of study in recent years. The
emerging picture now suggests that translational suppression through
eIF2 phosphorylation is an important component of
apoptotic programming (450). Thus, disruption of eIF2
phosphorylation may serve the dual purpose of
maintaining the translational competence of the host cell and
preventing apoptosis during host shutoff. This idea is
supported by the many studies of vaccinia virus replication in which
the viral K3L and E3L gene products have been implicated in disrupting
eIF2 phosphorylation and blocking apoptosis
(52, 55, 82, 83, 135, 255, 420). Moreover, studies by
Roizman and colleagues have demonstrated that disruption of eIF2
phosphorylation by the HSV-1 
134.5 gene
product was a requisite for sustained translational competence and
viral persistence during the host shutoff induced by HSV-1 infection
(188, 190). Influenza virus similarly ensures that eIF2
phosphorylation is blocked and translational competence
is maintained during host shutoff (244, 294). However, rather than preventing shutoff-induced apoptosis, influenza
virus may delay or reprogram apoptosis to facilitate cell lysis
and virion release during late-stage infection (116, 445,
446). In closing this section, it is important to note that
maintenance of translational competence during host shutoff may
ultimately contribute of viral pathogenesis. As described in "Viral
persistence and translational control" (below), the ability to
suppress mRNA translation is a key component for the control of
cell growth. In persistent viral infections, such as those by hepatitis
C virus (HCV) or the DNA tumor viruses, constitutive modulation of host translational control pathways and release of translational suppression may make important contributions to viral oncogenesis (125, 134, 248, 444).
Mechanisms and Control of Viral mRNA Translation
Viruses utilize the canonical translation factors and machinery of
the host cell to facilitate completion of their translational programming. Figure 5 depicts various
means by which viruses implement their translational programming toward
the common end of synthesizing viral proteins and completing the
virus life cycle. Reflecting the nature of the virus-host
relationship itself, the host cell has evolved countermeasures that
impose blockades upon viral protein synthesis. As described
below, viral translational programming often includes mechanisms to
manipulate the host translational machinery and overcome these
antiviral blockades.
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Internal ribosome entry.
Translation initiation,
mediated through the internal entry of ribosomes onto the substrate
mRNA, was first found in 1988 during studies of poliovirus and EMCV
replication (227, 370). Examination of the nucleotide
sequence of the picornavirus 5' UTR has revealed a
region of significant secondary structure spanning approximately 500 nucleotides (nt) and punctuated by multiple AUG codons (222,
228). This region was initially known as the ribosome landing pad
and later termed the internal ribosome entry site (IRES) (for detailed
reviews of the IRES, see references 193, 223, and
329). Translation studies performed in vitro and in
vivo demonstrated that the IRES could confer internal ribosome entry to
a downstream ORF when placed between the cistrons of a multicistronic
mRNA (226, 227, 370). Moreover, incorporation of the
IRES to precede the ORF of a circular mRNA facilitated ribosome
entry and translation of the circular cistron (57). These
studies concluded that the IRES is a genetic element that facilitates
internal ribosome entry and mRNA translation independent of the
m7cap structure. Since then, the observation of
IRES-mediated translation has been extended to include other virus
families, most notably the other picornaviruses and the
members of the genera Pestivirus and Hepacivirus
of the family Flaviviridae (which include bovine diarrhea
virus and HCV, respectively [Table 2])
(223). We also note that IRES-mediated translation of
certain cellular mRNAs has also been identified and may constitute
a minor proportion of total cellular mRNA translation within the
cell (223, 234, 235, 347).
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Ribosome shunt.
The processes of cap-mediated ribosome
entry, 5' scanning, and internal initiation are combined features of
the ribosome shunt mechanism of translation. Ribosome shunting allows
the small ribosomal subunit to avoid the problems of scanning a long
and complex linear sequence preceding the major ORF. Unlike
IRES-mediated translation, shunting is dependent on the 5' cap and the
cap-binding complex for ribosome entry onto the mRNA
(410) (Fig. 5). The best evidence for ribosome shunting, and
the general ribosome shunt model, comes from studies of the
pararetrovirus cauliflower mosaic virus (CaMV) (94, 410),
although shunting also takes place in other viral systems (reviewed in
reference 203). Upon engaging the mRNA, the
ribosome scans until it reaches a cis-acting shunting
element that promotes ribosomal translocation to a downstream
receiving element(s) (94). By this process, scanning becomes
nonlinear and the ribosome can bypass a large portion of the 5'
UTR to initiate translation at the downstream cistron (Fig. 5 and
7).
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Leaky scanning. Translation initiation site selection is determined, in part, by the context of the nucleotide sequence surrounding the first AUG codon encountered by the scanning ribosomal subunit. Departure from the canonical accAUGg sequence (the initiation codon is shown in bold letters) often results in the scanning ribosome initiating translation from this weak AUG at a low frequency or bypassing it completely in favor of a stronger downstream AUG start site (221). This AUG selectivity is referred to as leaky scanning. Leaky scanning allows the translation of multiple ORFs from a common mRNA substrate (Fig. 5). The versatility of leaky scanning is quite evident when one considers that each ORF need not be in the same reading frame. Thus, the process of leaky scanning allows the virus to maximize its genome coding capacity and encode functionally distinct proteins from a common mRNA.
Leaky scanning is widely used by viruses and is perhaps best defined from studies on retrovirus replication. Human immunodeficiency virus (HIV) encodes a heterogeneous class of mRNAs that include several multicistronic species. Among these are the bicistronic mRNAs encoding the viral Vpu and Env proteins (368). The ORFs for Vpu and Env are tandemly arranged such that the Vpu coding region precedes the Env ORF (Fig. 8A). Synthesis of Env is essential for viral replication and takes place through a mechanism of leaky scanning from the upstream Vpu ORF (368, 412). Env synthesis requires a weak Vpu translation initiation codon. Mutation of the weak Vpu start site to a sequence more closely matching the canonical start site sequence resulted in suppressed Env translation from the bicistronic Vpu-Env mRNA (412). Thus, the weak context of the Vpu initiation codon allows the ribosome to scan pass the Vpu ORF and to initiate translation at the downstream Env AUG codon (Fig. 8A). Analyses of HIV mutants in which the vpu gene was deleted or lacked the Vpu initiation codon revealed a stimulation of Env synthesis (413). Accordingly, vpu mutant viruses exhibited defects in virus release and showed increased syncytium formation in vitro (368). These results support a model for the coordinate expression of Vpu and Env during HIV infection, which is dependent on the presence of the vpu ORF. By this model, the synthesis of Vpu occurs inefficiently via a weak initiation codon, perhaps allowing Vpu to coordinately accumulate during infection to levels sufficient for its function in late-stage HIV replication. In accordance with the leaky-scanning mechanism, the synthesis of Vpu itself may coordinate Env production by impeding ribosome scanning to the downstream Env ORF. This model remains to be directly examined by analyzing the polyribosome distribution of the relevant mutant Vpu-Env mRNAs. However, it suggests that the limitations placed on Env translation by the upstream vpu ORF may allow Env expression to coincide with late-stage replication events, including virion assembly, and release. Thus, while ensuring that both viral proteins will be produced during HIV infection, translational control by leaky scanning provides for the coordinate expression of Vpu and Env during viral replication.
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Frameshifting.
As in the example provided by the HIV
genome, retroviral genomes exhibit overlapping gene arrangements (Fig.
8A). The mouse mammary tumor virus (MMTV) genome exhibits an
overlapping gag, pro, and pol gene
arrangement (72). Translation of these gene products
involves the process of frameshifting, in which the translating ribosome shifts position by +1 or 
1 nt, resulting in a change of
reading frame (reviewed by Gesteland and Atkins [151])
(Fig. 5). The process of frameshifting was first described from studies of Rous sarcoma virus replication (220) and has been
extensively defined in the MMTV and infectious bronchitis virus (IBV)
(a coronavirus)] systems, although it occurs widely throughout other
eukaryotic RNA viruses (151). Frameshift sites within the
viral mRNA correspond to heptanucleotide sequences in which the
mRNA slips 1 base with respect to the tRNAs in the A and P sites on
the translating ribosome (40, 108). The frameshift site,
also known as the slippery site, allows the tRNA to move along the
mRNA template by 1 base (forward or back) and reestablish
codon-anticodon pairing, resulting in a stable +1 or 1 reading frame shift.
1
reading frame. The heptanucleotide sequence that comprises the slippery
site typically conforms to the motif XXXYYYN. Frameshifting
occurs at this site through the slipping of two ribosome-bound tRNAs
that are translocated from the current reading frame of
X-XXY-YYN, to the 1 reading frame of XXX-YYY. In MMTV, translation initiation of the gag-pro mRNA begins at the
5' end of the gag gene (Fig. 8B). The translating ribosomes
encounter a slippery site and a pseudoknot structure near the
3' end of the gag gene. The majority of the ribosomes read
through this region, but approximately 25% hesitate at the
heptanucleotide site, where the mRNA will slip backward by 1 nt. This event is stabilized by the new pairing with the two tRNAs
in the 1 reading frame. Meanwhile, most of the translating
ribosomes will terminate to make Gag-Pro but another 10% will slip
again at the pro-pol site to make the requisite Gag-Pro-Pol
polyprotein (108).
What are the molecular mechanisms by which the tRNAs and
pseudoknot contribute to frameshifting during mRNA
translation? Evidence has accumulated to indicate that the actual
frameshift occurs at the second (underlined) codon of the tandem
slippery codon pair, XXXYYYN, corresponding to
the ribosome aminoacyl (A) site. Slippery A sites within
eukaryotic viruses correspond to the codon sequence of AAC, AAU,
UUA, UUC, and UUU (108). Interestingly, these codons
are decoded by tRNAs with a highly modified base in the anticodon loop
(185). Thus, it has been suggested that hypomodified
variants of these tRNAs may function to promote shifting by being less
bulky and therefore more easily moved within the slippery site
(186). However, this idea remains controversial, since other
researchers have proposed that frameshifting is mediated by standard
cellular tRNAs and is simply dependent on the strength of the
codon-anticodon tRNA interaction (40, 465). In either case,
frameshifting requires a pseudoknot structure near the slippery site to stimulate the frameshifting events.
Recent evidence indicates that the actual secondary structure of
the pseudoknot is important for stimulating
frameshifting. Analysis of the IBV frameshifting signals clearly
demonstrated that the pseudoknot causes ribosome
pausing. Replacement of the IBV pseudoknot with a simple
stem-loop structure of equivalent base pairs did induce ribosome
pausing but, remarkably, did not stimulate frameshifting
(427). These results suggested that ribosome pausing was
necessary but not sufficient for frameshifting to occur and support the
hypotheses that (i) conservation of pseudoknot structure is
essential for frameshifting and (ii) pseudoknot interactions with specific trans-acting factors may promote the
frameshift events. Atomic modeling of the MMTV gag-pro
pseudoknot supports the former hypothesis, in that this
pseudoknot does not have coaxially stacked helices but, rather,
assumes a wedge conformation induced by an A nucleotide between
the helices (418). Structure-function analyses of the MMTV
pseudoknot revealed that this A nucleotide was essential for
stimulating frameshifting activity. Structural analyses of other viral
pseudoknots should provide further insight into the
contribution of pseudoknot sequence and structure in ribosome frameshifting.
Control of termination and reinitiation.
Translational
control by reinitiation involves two or more tandemly arranged ORFs on
a common mRNA. In the simplest model of reinitiation, a short uORF
controls the translation of the major downstream ORF by impeding
ribosome scanning (reviewed by Geballe [146]). In this
sense, the uORF commonly asserts a suppressive effect upon translation
of the downstream ORF. However, and as in the CaMV 35S RNA translation,
exceptions to this rule do apply. As described above, the sORFA uORF of
CaMV actually plays a stimulatory role in translation of downstream
ORFs within the 35S RNA (195). An example of viral use of
reinitiation to negatively control translation from a downstream
cistron comes from studies of cytomegalovirus replication. During
cytomegalovirus infection, expression of the gp48 product of the
polycistronic viral gpUL4 mRNA is coordinately controlled to
reach peak levels during late-stage viral replication. gp48 is
translated from the third of three cistrons within the gpUL4 mRNA
(146, 313). Gelballe and colleagues have
determined that coordinate control of gp48 expression is mediated
through the actions of the second gpUL4 uORF (uORF2) (147).
Remarkably, the uORF2 inhibitory effect on gp48 translation is
dependent upon the sequence of uORF2 (48, 85); introduction
of uORF2 missense mutations severely diminished the inhibitory signal
upon gp48 translation, while introduction of mutations that preserved
the coding content of uORF2 led to retention of gp48 translational inhibition. In vitro and in vivo expression studies revealed that the
translational control actions of uORF2 (i) function exclusively in
cis to repress gp48 synthesis through ribosome stalling at the uORF2 termination codon and (ii) are mediated through interference of uORF2 translation termination by the uORF2 peptide product itself.
Analysis of uORF2 translation revealed that the 20-kDa peptide product
remained bound to the ribosome complex as a peptidyl-tRNA covalently linked to tRNApro, which decodes the uORF2
carboxyl-terminal codon (49). Recent studies have now
demonstrated that the uORF2 peptidyl-tRNApro blocks its own
hydrolysis and ribosome release to remain stably bound to the ribosomal
complex (50). These results suggest that inhibition of uORF2
peptidyl-tRNApro hydrolysis blocks the translation of gp48
by creating a barrier that obstructs ribosome scanning to the
downstream gp48 ORF (Fig. 9).
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1 capsid protein and the 1 nonstructural protein
(409, 417). Initial analyses of S1 mRNA translation revealed that it was translated inefficiently compared to other reovirus mRNAs (345). Subsequent in vitro studies showed
that ribosomes paused at several positions on the S1 mRNA relative to the S4 mRNA, suggesting that translating ribosomes were less evenly distributed along the coding region of the inefficiently translated S1 mRNA than of the efficiently translated S4 mRNA (97). These results were supported by in vivo studies in
which the distribution of translating ribosomes on polyribosome-bound reovirus S1 and S4 mRNAs was examined in reovirus-infected cells (98). The pattern of ribosome pausing in vivo showed that
ribosomes were less evenly distributed along the poorly translated
bicistronic S1 mRNA. Consistent with a model of S1 mRNA
translational control by reinitiation, expression of the downstream S1
ORF was significantly increased by mutation of the upstream AUG codon
to a less favorable context for translation initiation
(106). Interestingly, however, the synthesis of the upstream
S1 mRNA translation product was not decreased by the same
mutations. Identification of differential codon usage between the S1
ORFs suggests that the translation efficiency of the S1 uORF may be due
to codon usage that confers a low elongation rate through the
utilization of low-abundance tRNAs (106). The diminished
elongation rate of the S1 uORF may then limit the efficiency of
reinitiation and synthesis of the downstream S1 cistron. Studies aimed
at understanding the influence of differential codon usage on the
elongation rate may uncover additional examples of this type of
translation control among eukaryotic viruses.
As described in the examples cited above, control of reinitiation
has been attributed largely to processes inherent within the
5' UTR of the viral mRNA. Analyses of alfalfa mosaic virus (AMV) replication now suggests that the 3' UTR may likewise play an important role in translation reinitiation and the efficiency of viral protein synthesis. The single-stranded RNA genome of AMV is
capped but not polyadenylated. Translation studies have revealed that
AMV RNAs are efficiently translated in spite of lacking the traditional
poly(A) tail and the advantages to RNA stability and translation
afforded to polyadenylated transcripts (157, 200). In
contrast to the many viruses that induce host translational shutoff
during infection, AMV infection, AMV infection is not associated with a
decrease in host protein synthesis (149). How, then, do AMV
mRNAs adequately compete for available translation factors? Early
evidence suggested that the 3' UTR of the AMV coat protein played a
stimulatory role in mediating coat protein synthesis (399,
496). Examination of coat protein mRNA translation in vitro
and in vivo revealed this mRNA to be efficiently translated even in
the presence of large quantities of a cellular mRNA competitor. A
functional role for the 3' UTR in coat protein mRNA translation was
demonstrated by conducting similar experiments with mutant mRNAs
lacking the 3' UTR; loss of the 3' UTR consistently reduced the
efficiency of coat protein synthesis without altering mRNA stability (177, 399). Interestingly, it was found that the
3' UTR was required for assembly of the coat protein mRNA into
polyribosome complexes, indicating that the 3' UTR was an important
determinant for ribosome binding (177). Mutagenesis studies
were used to identity the 3' UTR nucleotide sequence element GAUG as an
important determinant in AMV coat protein synthesis. This
tetranucleotide sequence encompasses an initiation codon
downstream from the coat protein termination codon and is thought to
stimulate coat protein synthesis through a process of
reinitiation (177). With this model, reinitiation would
facilitate ribosome-mRNA interaction and continued coat protein synthesis.
How might reinitiation within the 3' UTR actually contribute to
increased translational efficiency? One possibility is that reinitiation may retain the mRNA within the pool of active
ribosomes, thereby increasing the probability of 5' UTR-ribosome
interactions and promoting further rounds of authentic translation
initiation. On the other hand, the viral 3' UTR may stimulate coat
protein translation through a process independent of reinitiation,
although this idea remains inconsistent with experimental evidence. In this case, it remains possible that specific 3' UTR-protein
interactions may impart increased translational efficiency.
Functional recoding. In addition to frameshifting, viruses partake in functional recoding whereby translation proceeds through an in-frame termination codon (Fig. 5). This occurs though a process of redefining the termination codon to encode glutamine at UAG or tryptophan or selenocysteine at UGA (151). Functional recoding has been extensively studied in the Moloney murine leukemia virus (MuLV) system. MuLV redefines the stop codon at its gag-pol junction through the insertion of glutamine at the UAG stop codon. This allows the translating ribosomal complex to read through the gag-pol junction and synthesize the Gag-Pol polyprotein. During viral replication, the ribosome reads through the MuLV gag-pol stop codon 5% of the time, and this exact frequency seems to be essential for replication (491). If the UAG codon is replaced with an in-frame GAG codon, no viral particles are formed (111). In contrast, replacement of the native gag-pol stop codon with either the UAA or UGA stop codon permitted translational readthrough with similar efficiency. These results suggest that MuLV utilizes functional recoding as a mechanism to control the level of Gag-Pol polyprotein synthesis during the course of infection.
Moreover, it appears that redefining the stop codon is not codon specific, suggesting that other elements within the viral mRNA are responsible for translational readthrough. It is now clear that sequences downstream of the stop codon are necessary for functional recoding. In MuLV, this includes a pseudoknot sequence that appears to stimulate the recoding process (112, 482) (Fig. 10). Structure-function analyses of the MuLV pseudoknot revealed several interesting features required for stimulating recoding, including (i) specific nucleotide sequence within the spacer region between the stop codon and the pseudoknot; (ii) nucleotide conservation within stem-loop 2 of the pseudoknot; and (iii) a nonhelical pseudoknot structure (482). These results support the idea that recoding may be dependent upon recruitment of trans-acting factors to the termination site, possibly mediated through sequences within the spacer region between the stop codon and the pseudoknot and/or the pseudoknot structure itself. Identification of such factors may have implications for the development of future antiviral drugs, since disruption of the recoding process results in a block in virion formation within the infected cell (111).
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Coupling the Virus Life Cycle to Translational Control
Translation control programs in eukaryotic cells play important roles in governing cellular metabolism. In many cases, these programs are implemented in response to specific environmental cues. Viruses, specifically the more complex DNA viruses, including the poxviruses, papillomaviruses, and herpesviruses, have similarly incorporated into their own life cycles translational control programs that play important roles in replication, latency, and virulence. In particular, recent evidence from studies of HSV replication indicates that translational control programs are implemented by the virus to (i) facilitate host shutoff, (ii) control global and specific viral gene expression, (iii) maintain or exit latency, and (iv) overcome the host antiviral response. In this section, we use the herpesvirus life cycle to illustrate how viruses utilize translational control programs to direct replicative decisions and how this translational programming contributes to the control of viral replication.
The herpesviruses. The human herpesviruses establish latent infection in either neural cells (HSV and varicella-zoster virus) or hematopoietic cells (Epstein-Barr virus and cytomegalovirus) with negligible damage to their respective host. Latency permits viral persistence in the face of an active immune response. In response to certain stimuli, the viruses may periodically reactivate from latency throughout the life of the host to enter the productive phase viral life cycle, which sheds sufficient virus progeny to infect new hosts. This life-style is bound to impose a distinctive set of evolutionary pressures on the control mechanisms regulating herpesviruses gene expression. Thus, herpesviruses present a challenging and attractive model system for studying the ever-complex mechanisms of viral gene expression and regulation at the level of translation. Furthermore, mutant herpesviruses deficient in a particular function can be isolated or genetically engineered with deletions in certain genes to assess the roles of specific virus-encoded proteins in viral translation and replication in the host. In the following sections, we examine how the alphaherpesviruses HSV-1 and HSV-2 have implemented control of viral and host translation into their latent and productive phases and discuss how translational control of HSV mRNA might contribute to viral pathogenesis.
Because HSV gene expression is coordinately modulated during a productive infection, the viral genes can be categorized into three kinetic classes: immediate-early (), early (
), and late ()
genes. genes are transcribed in the absence of de novo protein synthesis; this process peaks at 2 to 4 h postinfection, and the transcripts continue to accumulate until late in infection. Most products of genes are potent transcriptional
trans-activators that cooperate to activate the
transcription of and genes. genes encode proteins
required for HSV DNA synthesis, as well as a number of auxiliary
replication factors. Viral structural proteins are the products of genes, whose expression occurs at the onset of DNA synthesis. While
much research on HSV has centered on the transcriptional events
responsible for the differential expression of , , and genes,
accumulating evidence indicate that translational mechanisms are also
important for HSV gene expression.
HSV and the shutoff of host cell protein synthesis. Like other cytolytic viruses, HSVs are thought to facilitate their replication by preferentially producing viral proteins at the expense of host cell gene expression. In tissue culture cells infected with HSV-1 or HSV-2, host protein synthesis and mRNA levels decrease by approximately 90% within 3 h postinfection and viral proteins dominate thereafter (424). This remarkable feature of the shutoff of host protein synthesis induced by HSV infection, presumably to alleviate competition for precursors, is a multistep process that involves several mechanisms and can be separated into two stages: primary shutoff and secondary shutoff. Primary shutoff, which is characterized by rapid disintegration of preexisting polyribosomes and degradation of preexisting cellular and viral mRNAs, occurs very early after HSV infection in the absence of de novo protein synthesis. In contrast, secondary shutoff takes place later in the course of infection and requires viral gene expression.
Despite extensive research efforts, the exact mechanisms responsible for the shutoff events during HSV-1 infection are poorly understood, but encouraging progress has been made over the past few years. In the primary shutoff, at least one viral factor, the virion host shutoff (VHS) protein, is necessary for mRNA destabilization, and this may, at least in part, account for the disassociation of polyribosomes (280). Encoded by HSV gene UL41, the VHS protein is a 58-kDa phosphoprotein located between the capsid and envelope regions of the virion (called tegument) and is delivered into the cytoplasm of newly infected cells. How does VHS function to specifically induce mRNA degradation? Apart from having limited homology to a small segment of PABP (389), the primary sequence of the VHS protein provides little clue to its function. Despite the lack of any primary sequence similarity to known RNases, several lines of evidence suggest that VHS is associated with RNase activity: (i) incubation of polyribosomes from uninfected cells with postpolysomal (S130) supernatant from HSV-infected cells, but not S130 from uninfected cells or from cells infected with a VHS-defective virus, resulted in rapid degradation of stable mRNAs; (ii) crude extracts from host shutoff-competent virions or reticulocyte lysate containing wild-type VHS protein displayed enhanced RNase activity, while VHS mutants did not; and (iii) antibodies against VHS protein inhibited the RNase activity of wild-type VHS protein in the cell-free reactions. However, there is as yet no direct evidence, obtained using highly purified proteins, to demonstrate that wild-type, but not mutant, VHS protein itself indeed contains RNase activity in vitro. Thus, the possibility that the VHS protein works in conjunction with another factor(s) with RNase activity has not been entirely excluded. The mechanism(s) by which the VHS protein specifically targets mRNA for degradation is not known. Polyadenylated RNAs are degraded faster than nonpolyadenylated substrates (225), and deadenylated mRNAs congregate in HSV-infected cells (180). Thus, it is conceivable that the VHS protein recognizes polyadenylated mRNAs, perhaps through the putative poly(A)-binding region of VHS. In this context, it would be interesting to test the effect of mutating the conserved residues in this region on HSV-induced mRNA degradation and translational arrest. Furthermore, the ability of the VHS protein to bind poly(A) mRNA in vivo (for example, through UV cross-linking procedures) or the RNA sequences or structural regions recognized by VHS have not been determined. Alternatively, the VHS protein may interact with the PABP complex for localized poly(A) cleavage of mRNA. This possibility is consistent with the observation that mRNAs in infected cells or in cell-free reaction mixtures are preferentially degraded over protein-free RNAs, suggesting that the VHS protein might achieve specificity through interaction with a factor(s) present in the ribonucleoprotein (RNP) complex. Therefore, the identification of VHS-interacting factors may also provide insights into the mechanistic action of the VHS protein. At any rate, considering the specificity of the VHS protein and the generation of discrete decay mRNA intermediates, VHS probably operates, either on its own or in cooperation with another factor(s), by cleaving mRNA molecules at one or a few critical sites, such as poly(A), that normally function to protect mRNA from destruction by host RNases. The ICP27 protein of HSV-1 is thought to play an important role in the secondary shutoff of host protein synthesis (180). A nuclear phosphoprotein of 63 kDa, ICP27 was originally known for its capability to both stimulate and repress the expression of different target genes. Subsequent studies demonstrated that ICP27 is also capable of interfering with host cell splicing, causing a reduction in the levels of several cellular spliced transcripts and an accumulation of pre-mRNA in the nucleus during infection (180, 181). Furthermore, ICP27 expression causes a dramatic redistribution of the splicing small nuclear RNPs and other splicing factors during HSV-1 infection (406). Finally, an interaction between ICP27 and the splicing machinery has been demonstrated (181, 406). On the basis of these studies, it has been hypothesized that ICP27-mediated impairment of host cell splicing may contribute to the secondary shutoff, because unspliced cellular transcripts remain in the nucleus, where they become degraded. This leads to fewer cellular transcripts being exported to the cytoplasm for translation, and thus selective synthesis of virus-specified proteins is favored, since most viral transcripts are not spliced.Selective repression of mRNA translation initiation during HSV infection. HSV induces shutoff of most host protein synthesis. However, a few remaining cellular mRNAs, whose protein products presumably play an important role in the survival of the virus within the host, are continuously being translated after infection (285). This paradox raises some important questions. (i) What is the function and nature of these cellular mRNAs? (ii) Is the sustained translation of cellular mRNAs induced by HSV or an inherent feature of the mRNAs? (iii) What is the mechanism of the persistence of translation of these mRNAs after infection?
Recent studies demonstrate that at least one set of cellular mRNAs that are persistently translated after HSV-1 infection encode ribosomal proteins (164, 425). An analysis of ribosomal protein mRNA expression across a polyribosome gradient revealed that there is a discernible shift between the untranslated subpolysomal (prepolysomal) fraction to the polyribosomal fraction as infection proceeded (164). It is known that the 5' leader sequence of vertebrate ribosomal protein mRNAs contains a terminal oligopyrimidine tract (known as the 5' TOP element) that is sufficient to confer translational regulation and migration between the polyribosomal fractions (13). The specific recruitment of 5' TOP mRNAs by ribosomes is closely associated with an increase in phosphorylation of ribosomal protein S6 (231, 232, 456). In this regard, S6 proteins in preexisting ribosomes were phosphorylated to greater extent than were those found in newly assembled ribosomes in infected cells (164). This suggests that translation of ribosomal protein mRNAs may occur preferentially on preexisting ribosomes. Interestingly, in parallel with this study, a progressive shift of-actin and GAPDH
mRNAs from polyribosomes to 40S subunits was observed during the
course of infection. This phenomenon appeared to be independent
of VHS-mediated mRNA degradation, since the protein did not affect
mRNA recruitment by polyribosomes. With the caveat that only two
cellular mRNAs have been examined, these studies suggest that
HSV-1 may employ an additional strategy to selectively suppresses host
mRNA translation, namely at the initiation step.
Other mechanisms may also contribute to the persistence of ribosomal
protein synthesis. In addition to S6 ribosomal protein, HSV-1 infection
induces phosphorylation of at least two other proteins,
including the product of the US11 late gene (90). Furthermore, the possibility that an increase in the elongation rate of
translation might also account for the sustained translation of
ribosomal protein mRNAs has not been eliminated. In this regard, two HSV-1 proteins, the trans-activator ICP0 protein
(10) and the protein kinase encoded by the
U(L)13 gene (257), interact with and
phosphorylate elongation factor-1 delta (EF1-
), respectively. However, a role for EF1- phosphorylation in HSV-1
replication has not been demonstrated.
Selective translation of HSV mRNA. How does HSV achieve selective viral translation during the shutoff of protein synthesis? In VHS-induced shutoff, in which both cellular and viral mRNAs undergo concomitant degradation, HSV would need to ensure that the viral mRNAs continue to accumulate after cellular mRNAs have been degraded. The potency and indiscriminate nature of the VHS activity suggests that it would have to be negatively controlled in a temporal fashion during the course of infection. Evidence for this hypothesis came from a study demonstrating that the virion trans-activator VP16, which forms a specific complex with VHS in the infected cell, is capable of suppressing VHS activity (282). Specifically, viral protein synthesis and mRNA levels were significantly reduced at intermediate times after infection with a VP16 null mutant virus. Additionally, a stably transfected cell line expressing VP16 was refractory to VHS-induced host shutoff of protein synthesis. Although it remains to be shown, the VHS binding function of VP16 is likely to be important for inactivating VHS, either by masking one or more functional domains, inducing a conformational change, or by targeting VHS into the nucleus and/or the virion assembly pathway. Moreover, the mechanism by which the VP16-VHS interaction is modulated is not known. Finally, it is noteworthy that two other viral factors are involved in HSV-induced host shutoff: the virion-associated protein kinase encoded by the U(L)13 gene (358) and the ICP22 protein encoded by the US1.5 gene (349). The possibility that these gene products may mediate host shutoff by regulating VHS and/or VP16 function will undoubtedly be explored in forthcoming studies.
Another strategy by which HSV may exert selective translation of viral mRNAs over host mRNAs is suggested by recent studies that demonstrate a role of the ICP27 protein in the export of HSV-1 intronless mRNAs (405). Thus, ICP27 appears to mediate preferential viral translation via two mechanisms: (i) it impedes the translation of cellular mRNA by preventing the export of this mRNA to the cytoplasm through the impairment of host splicing, and (ii) it binds viral transcripts and delivers these RNAs to the cytoplasm for translation. However, several key aspects will have to be elucidated in future studies to strengthen the proposed dual role of ICP27. These include assessing (i) the functional significance (an effect on splicing) of the interactions and changes of ICP27 and small nuclear RNPs, (ii) the specificity of ICP27 RNA binding, and (iii) the interaction of ICP27 with the cellular nuclear export complex. Finally, the observations that ribosomal proteins are persistently synthesized and new ribosomes are assembled after HSV-1 infection might suggest another mechanism for the preferential translation of viral mRNAs. Because new ribosomes contain underphosphorylated S6 ribosomal protein, they are presumably less effective in the translation initiation of mRNAs that possess 5' TOP sequences (reviewed in reference 232). Thus, viral mRNAs which lack 5' TOP sequences may be selectively initiated by newly synthesized ribosomes over cellular mRNAs containing 5' TOP. Although the idea is not unreasonable, especially since reinitiation of translation progressively becomes a limiting factor during shutoff (10, 285), it remains to be supported by any experimental evidence.Inhibition of eIF2 phosphorylation
during HSV infection.
Cells modulate the synthesis of proteins in
response to external stimuli, including viral infection, through the
modification of translation factors. Phosphorylation of eIF2 is
perhaps the best-characterized mechanism by which this occurs,
particularly within the context of virus infection. As described in
detail in "Viral modification of translation factors" (below),
viral replication produces highly structured viral transcripts in the form of dsRNA that can bind to and activate the host PKR, which in turn
phosphorylates eIF2 (131). As a result, the cellular translational machinery is incapacitated and viral protein synthesis and replication are restricted within the infected cell. Accumulating evidence now indicate that HSV-1, like many viruses (see Table 4), has
evolved ways to circumvent the virally induced translational block by
counteracting PKR function (Fig. 11).
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134.5 gene. These viruses fail to grow on many human
malignant neuronal cells, which displayed increased PKR and eIF2
phosphorylation, as well as premature shutoff of
protein synthesis late in infection. The phenomenon appeared to be
independent of VHS function and mRNA degradation (62,
379). Interestingly, an unknown 90-kDa phosphoprotein (p90)
coprecipitated with anti-PKR antibody from lysates of cells infected
with 134.5
viruses. While the function of
p90 is not known, the correlation between p90
phosphorylation and the premature shutoff of protein synthesis suggests that it may play a positive role in modulating PKR
activity in phosphorylation of eIF2. It was thought
that the 134.5 gene product might inhibit protein kinase
activity by blocking the interaction between p90 and PKR. However,
subsequent studies demonstrated that the 134.5 protein
operates through a different mode of action. Using the yeast two-hybrid
approach, He et al. (190) found that the
134.5 protein associated with the cellular protein
phosphatase 1 (PP1). Further, the 134.5 protein
formed a complex with PP1 in HSV-1-infected cells, and fractions
containing the complex were capable of dephosphorylating purified
eIF2. Thus, the 134.5 protein is likely to function as a regulatory or targeting subunit of PP1 to redirect the
phosphatase to dephosphorylate eIF2, therefore neutralizing
PKR activity. However, it is not clear how the 134.5
protein guides PP1 to eIF2; an interaction between
134.5 and eIF2 has not been demonstrated. Interestingly, we have recently obtained evidence that PP1 can also
directly inhibit PKR function by binding to and dephosphorylating PKR
(S.-L. Tan and M. G. Katze, unpublished data). Whether
dephosphorylation of PKR by PP1 is also triggered by
HSV-1 infection or other signals is not known. Furthermore, it will be
important to identify the regulatory subunit of PP1 that targets the
phosphatase to PKR (Fig. 11).
The story becomes more complicated with recent studies describing the
isolation of second-site suppressor mutant viruses that lack the
134.5 gene (53, 338). These variant viruses
regained the ability to grow on otherwise nonpermissive neuronal cells and contained additional mutations that affect a distinct viral genetic
element, the SUP locus. Moreover, deletion of the SUP locus
prevented the accumulation of phosphorylated eIF2. Consequently, extragenic suppressor 134.5 mutants could sustain
protein synthesis and multiply on cells that failed to support
replication of the parental the 134.5 variants. As it
turns out, these dominant suppressor alleles compensated for the loss
of the 134.5 function by overproducing a viral
RNA-binding, ribosome-associated protein (US11) that reduced PKR
activation (343). Taken together, these results suggest that
HSV-1 encodes at least two strategies (US11 and 134.5)
to modulate cellular translation by targeting both PKR and eIF2
(Fig. 11). Interestingly, the 134.5 protein contains a
region of significant homology to the cellular protein GADD34, which is
induced in response to agents that promote growth arrest, DNA damage,
and differentiation. Indeed, GADD34 could also interact with PP1 and
could functionally replace 134.5 in prolonging late-protein synthesis in infected cells (189). These
studies suggest that signals that trigger differentiation, growth
arrest, and DNA damage may be intimately linked to translational control.
Implications of viral modulation of translation in HSV pathogenesis and disease treatment. An important question that has remained unanswered in the study of HSV pathogenesis concerns how the viruses undergo latency in their host. It is unanswered because of the lack of a reliable cell culture system to support latent HSV infection. What is known from in situ hybridization studies of latently infected neuronal cells is that the latent state is characterized by the transcription of specific colinear transcripts. These RNAs, known as latency-associated transcripts (LATs), persist despite the absence of virtually all gene expression. Although experiments with LAT-negative mutants showed that the LATs are not required for HSV-1 lytic replication and establishment or maintenance of latency, they appear to be necessary for efficient in vivo reactivation in infected animal models. Thus, there is great interest in determining the stage of latency at which the LATs modulate, their mechanisms of action, and the exact sequences responsible for reactivation. In this regard, Steiner and colleagues have recently reported that the LATs are associated with polyribosomes in vitro and during latent in vivo infection (158, 159). These observations are highly suggestive of translation of a functional LAT protein, although efforts to demonstrate the presence of HSV-1 LAT protein products in latently infected tissues have been futile to date. To begin to gain an understanding of how translational control may play a role in HSV reactivation and thus contribute to viral pathogenesis, it will be critical to identify mechanism(s) of ribosome binding to the LAT transcripts and to characterize the LAT proteins.
Finally, the cytolytic and neurotropic properties of HSV-1 render this virus a potential tumoricidal agent for destroying malignant cells in the central nervous system. In this regard, HSV-1134.5
mutant viruses, which are attenuated and nonneurovirulent in animals,
are able to discriminate between normal and malignant cells
(8). However, these variant viruses grow poorly on neuronal tumors, which imposes a major limitation on their effectiveness in
destroying tumor tissue. The suppressor mutants of the
134.5 allele, which have regained the ability to grow on
neoplastic cells but retained the attenuated phenotype of the
134.5 parent virus, may represent a prototype virus to
destroy malignant cells in the CNS. Furthermore, the use of attenuated
HSV as a vector for gene therapy in the study and treatment of
neurodegenerative diseases is under investigation.
Recruitment of Host Factors for the Efficient Translation of Viral mRNA
The previous examples of HSV-host interactions and the resulting modification of host factors reflects a common theme among viruses, which target and modify host process to facilitate viral replication. Indeed, the pressures of translational dependence on viral replication have resulted in a wide variety of viral strategies to maximize translational efficiency. Such strategies often involve recruitment of specific host factors that function to improve the efficiency or mediate the selectivity of viral mRNA translation. This section discusses how viruses recruit and utilize specific host factors, in addition to the conserved repertoire of canonical translation factors, to facilitate efficient mRNA translation during infection. As presented below, recruitment of host factors to the viral mRNA is broadly used in both cap-dependent and cap-independent translational strategies. This section will begin by examining the host factors that are recruited to the IRES element and the viral 3' UTR and discussing the roles that such proteins may play in mediating viral mRNA translation and mitigating virus host range. This is followed by a detailed examination of how influenza virus ensures the efficient and selective translation of its mRNAs though a process of host factor recruitment and modification.
IRES binding proteins and proteins that bind the viral 3' UTR. As the functional element for cap-independent translation initiation, the IRES promotes viral mRNA translation through the recruitment of canonical translation factors to the initiation site (223). In addition, it is now known that several noncanonical factors are recruited to the IRES that stimulate, or in some cases repress, IRES-mediated translation. Structure-function analyses have provided insights into the roles of IRES-binding proteins in viral replication and have identified sequences within the IRES that direct interaction specificity (162, 238, 239, 394) (reviewed by Belsham and Sonenberg [23], Jackson and Kaminski (223), Hellen and Wimmer [193], and Ehrenfeld [101]). Meanwhile, protein interactions with the viral 3' UTR have similarly been implicated in providing translational efficiency and selectivity during viral infection (122, 209, 218, 302, 360, 496). A role for 3' UTR-binding proteins in viral mRNA translation may reflect the efforts of viruses to take full advantage of the translational efficiency provided by the closed-loop translation model.
The early observations from in vitro studies revealed that efficient translation from enterovirus or rhinovirus IRES in rabbit reticulocyte lysate (RRL) required supplements derived from HeLa cell extracts (41, 99). In contrast, the cardiovirus-aphthovirus IRES conferred efficient translation in native RRL. These results were significant in that they indicated that IRES translation required a specific trans-acting factor(s) supplied by the host cell, pointing the way to the identification of IRES-binding proteins. Moreover, these observations implicated IRES-binding proteins in the determination of virus host range specificity. IRES-binding proteins have since been identified through a combination of gel mobility shift assays and the use of UV light to cross-link RNA probes with resident cytoplasmic factors within cell extracts. Several distinct cellular IRES-binding proteins have been identified and characterized (reviewed in reference 23). Among the best-characterized IRES-binding proteins are the systemic lupus erythematosus autoantigen (La) and PTB (36, 191, 194, 328, 330). Both proteins specifically bind the poliovirus IRES. La is a 52-kDa RNA-binding protein that is known to interact with RNA polymerase III-transcripts and to play a role in the transcription termination reaction (160). The La protein resides predominantly within the nucleus of uninfected cells but, importantly, is redistributed to the cytoplasm during picornavirus infection (330). Structure-function analyses have identified nucleotides 559 to 624 of the poliovirus RNA as the major binding site for the La protein (328). This region maps to within the polypyrimidine tract of the poliovirus IRES (Fig. 6). The best evidence in support of a functional role for the La protein in poliovirus translation comes from experiments in which RRL was supplemented with purified recombinant La protein. Addition of recombinant La stimulated poliovirus translation to an efficiency similar to that observed when RRL was supplemented with HeLa extract (330). However, the concentration of recombinant La greatly exceeded the level of La within HeLa extracts, suggesting that (i) recombinant La was only partially active or (ii) additional factors may participate with La to promote poliovirus translation. It should be noted that the La protein also binds to the 5' UTR of influenza virus and HIV RNA, where it may also play roles in facilitating efficient viral protein synthesis (361, 443). Thus, the functional role of La in viral protein synthesis may not be limited to promoting IRES-mediated translation but may reflect a more general function. Although a direct biochemical role for La in IRES-mediated translation remains to be demonstrated, it is postulated that La may function as an RNA chaperone to maintain RNA structure in a conformation that favors translation. PTB is a 57-kDa cellular protein that plays a role in RNA splicing (304). PTB binds to multiple sites within the poliovirus IRES (191), but a direct role for PTB function in poliovirus translation has yet to be demonstrated. PTB also binds with high affinity to the EMCV IRES (229) and, more recently, to the IRES of HCV (3, 4, 217). Major PTB-binding sites have been identified within stem-loop H and the polypyrimidine tract of EMCV (229) (Fig. 6). Similar to the La protein, PTB is postulated to function as an RNA chaperone, where it may stabilize IRES structure in a translation-competent conformation. Evidence to support this idea comes from analyses of EMCV RNA that possessed point mutations within the stem-loop H PTB-binding site (229). Mutations that disrupted stem-loop base pairing prevented PTB binding and abrogated IRES function. Meanwhile, compensatory mutations that restored the native stem-loop conformation restored both PTB-binding and IRES function. A direct role for PTB in EMCV IRES-mediated translation was suggested from competition experiments in which PTB was functionally depleted from in vitro translation reaction mixtures by the addition of competitor RNA that contained a PTB-binding site (38). Addition of competitor RNA was found to specifically inhibit EMCV translation, and the addition of exogenous PTB relieved this competition to restore EMCV translation. However, recent results from Kaminski and Jackson (239) suggest that PTB binding may not be an absolute requirement for EMCV translation but, rather, is conditional upon the structure of the major PTB binding site within the EMCV IRES. In addition to La and PTB, several other factors have been identified that may play a role in IRES-mediated translation, although the nature of such factors remain to be determined (23). Recent analyses of HAV IRES function indicate a role for PCBP2 in HAV translation (162). PCBP2 was found to bind specifically to within the polypyrimidine tract of the HAV IRES, corresponding to HAV nt 1 to 157. In these experiments, affinity column depletion of PCBP2 from HeLa extracts resulted in only low levels of HAV translation while translation was restored by adding back recombinant PCBP2 to the depleted extracts. PCBP2 has also been implicated in stimulating poliovirus translation (35), where it has been shown to bind to stem-loop IV of the poliovirus IRES (34). An essential role for PCBP2 in HAV and poliovirus translation awaits further studies. However, like La and PTB, it appears that PCBP2 may play a role in maintaining IRES structure in a translationally competent conformation. HAV RNAs lacking the 5'-terminal 138 nt, which are not part of the HAV IRES but do include a major region of the PCBP2-binding site, retained translational efficiency independent of PCBP2 (162). This suggests that deletion of the first 138 nt of the HAV RNA may mimic the effects of PCBP2 binding and allow the IRES to spontaneously adopt the correct structure needed to promote translation. Considering that the picornavirus genomic RNA must serve as template for both translation and transcription, one may propose that at least one role for IRES-binding proteins like La, PTB, and PCBP2 is to functionally differentiate between translation and transcription by making the RNA accessible to the translational machinery. By this model, these and other proteins may bind to their cognate motif(s) within the IRES to induce and/or stabilize the specific RNA tertiary structures that promote mRNA translation over genome transcription. On the other hand, modification, or masking, of IRES-binding proteins by viral and/or cellular factors may provide the signals promoting the switch from translation to transcription. Recent studies indicate that the IRES and IRES-binding proteins may not act alone to promote cap-independent viral mRNA translation. In addition to an IRES, the HCV genome contains a highly structured 98-nt 3' UTR (32, 265). Sequence analyses revealed that the HCV 3' UTR is highly conserved among viral isolates, suggesting that it may function as a requisite element in HCV replication. This region of the HCV genome contains a high-affinity PTB-binding site (216, 464). Interestingly, Lai and colleagues found that the presence of this PTB-binding site actually relieved translational repression conferred by PTB binding to a second high-affinity site located within the HCV IRES (217, 218). It has been proposed that PTB may control IRES-mediated translation by interacting with both the viral RNA and an unknown factor(s) to (i) bind the viral 3' UTR and relieve translational repression from within the HCV IRES and (ii) enhance translation through circularization of the HCV RNA into a closed-loop translation complex. The exact role of PTB-mediated translational suppression and stimulation in the context of HCV replication have yet to be determined. One possibility is that PTB interactions with discrete regions of the HCV genome ensure the efficient translation of only genome-length RNA, and provide translational fidelity to HCV polyprotein synthesis. Another 3' UTR-directed mechanism of stimulating viral mRNA translation has been described in rotavirus-infected cells. In contrast to cellular mRNAs, rotavirus mRNAs lack a poly(A) tail. Viral mRNA stability is achieved in part through the actions of the viral NSP3A protein, which binds to the 3' end of virus-encoded mRNAs. During rotavirus infection, NSP3A directly interacts with the eIF4G isoform, eIF4GI (376). Interestingly, NSP3A may represent a virus-specific analog of PABP, in that it appears to compete with PABP for interaction with eIF4G and participation in the translation initiation process. Results from in vitro experiments suggest that NSP3A may actually "evict" PABP from the eIF4F translation initiation complex. These results support a model in which NSP3A mediates selective translation of viral mRNA by (i) disrupting the cellular mRNA translation through "eviction" of PABP from the "closed-loop" translation complex and (ii) facilitating viral mRNA translation through interactions with eIF4G and the viral mRNA 3' end. Moreover, eviction of PABP from the translation complex is likely to contribute to host protein synthesis shutoff during rotavirus infection by reducing the overall efficiency of host mRNA translation.Influenza virus. Similar to the previous examples provided by the picornaviruses, influenza virus depends on the recruitment of specific host factors to mediate selective and efficient translation of viral mRNAs during infection. Influenza virus is responsible for up to 70,000 deaths a year in the United States and 20,000,000 worldwide in the worst epidemic years and remains one of the most dreadful threats for a recurring virus pandemic. Like other cytopathic viruses, influenza virus dramatically perturbs the normal synthesis of host macromolecules. However, unlike most nononcogenic RNA viruses, the replicative cycle of influenza virus includes both a nuclear and cytoplasmic phases. Accordingly, the evolution of the translational strategies of this virus is likely to be dictated by a different set of selective pressures (274). Given its small negative-strand, segmented RNA genome size, it is almost intuitive that such a successful virus must have evolved a plethora of ingenious schemes to ensure selective and efficient translation of viral mRNAs. In the sections that immediately follow, we review our present understanding of how influenza virus imposes selective translation of viral mRNAs over cellular mRNAs, but yet manages to keep the infected cells translationally competent, such that the steps involved in the protein synthetic pathway are not compromised.
Selective translation of influenza virus mRNAs.
The mechanisms by which influenza virus mediates selective
translation of viral mRNAs are summarized in Table
3. In influenza virus-infected cells,
cellular mRNAs are subjected to a modest decrease in transcription
rates (approximately twofold) (250, 251). In addition,
degradation of cellular mRNAs is evident late after infection
(22, 215). Moreover, newly synthesized cellular mRNAs
fail to reach the cytoplasm after infection due the degradation of
nuclear RNA early after infection (250). Although the
mechanism has not been determined, it is thought that influenza
virus-induced cellular mRNA degradation in the nucleus is initiated
by the cleavage of the 5' ends of cellular RNA polymerase II
transcripts by the viral cap-dependent endonuclease. The decapped
RNAs would probably be more susceptible to degradation by cellular
nucleases, since it is well established that the 5' cap structure
stabilizes mRNAs (274).
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Contribution of influenza virus mRNA structure upon selective translation. It was conceivable that translational selectivity, at least at the level of initiation, could be due to competition between cellular and viral mRNAs for limiting components of the translational machinery as shown in other systems, including the reoviruses (309, 388, 471). Since viral mRNAs do not have an advantage merely due to sheer mass (143), it was likely that the influenza virus mRNAs are intrinsically better initiators of translation due to certain unknown structural qualities. Such structural features could deceive the cellular protein-synthesizing machinery into making only viral proteins (141, 143). The first persuasive evidence that influenza virus mRNAs are intrinsically efficient initiators of translation was provided by studies in which cells were doubly infected with influenza virus and adenovirus (246, 247, 249). These early experiments also showed that influenza virus also has a strategy to sustain overall high levels of protein synthesis.
In these adenovirus-influenza virus doubly infected cells, influenza virus proteins accumulated essentially to the same levels as in cells infected by influenza virus alone (246). These data suggested that influenza virus is able to overcome the halts on host cell mRNA transport and translation imposed by adenovirus (14), demonstrating that the virus establishes its own translational and transport regulatory mechanisms. Furthermore, influenza virus mRNAs were more efficiently translated than late adenovirus mRNAs, which are believed to be strong mRNAs due to the presence of the tripartite leader sequences (310). Further evidence that the structure of influenza virus mRNAs plays a key role in their selective translation was obtained from a study conducted by Alonso-Caplen et al. (5). The authors found that the translation rate of the influenza virus nucleocapsid protein (NP) mRNA, expressed from a recombinant adenovirus, was equivalent to that of the native NP expressed by influenza virus itself. The recombinant NP mRNA translation rates were remarkably efficient despite the absence of all other influenza virus gene products. Thus, these studies indicated that the sequence and/or structure alone of an influenza virus RNA can confer enhanced translatability. Because influenza virus is not readily amenable to genetic studies, researchers have resorted to alternative strategies to study the contribution of the viral mRNA structure to selective translation. Definitive evidence that influenza virus mRNAs have an innate ability to be preferentially translated was obtained from transfection and infection studies in which representative viral or cellular cDNAs were transfected into COS-1 cells, which were then infected with influenza virus (140). It was shown that mRNA translation, directed by cellular transfected genes such as interleukin-2 or secreted embryonic alkaline phosphatase (SEAP) was markedly shut off after viral infection. In contrast, an exogenously introduced influenza virus gene encoding NP was not subjected to the translational blocks imposed on the cellular genes. Subsequent studies using chimeric constructs in which the viral 5' UTR was fused to a cellular mRNA demonstrated that the selective translation of influenza virus mRNAs is mediated almost exclusively by the viral 5' UTR (142).Influenza virus recruitment of Grsf-1.
Unlike the IRES
of poliovirus or the tripartite leader of adenovirus, influenza virus
mRNAs do not possess any extensive secondary structures. Instead,
they contain short and relaxed 5' leaders with no upstream AUGs. Using
gel mobility shift and UV cross-linking analysis, several factors have
been identified that bind to cis-acting sequences present in
the viral 5' UTR (361). A yeast three-hybrid screen using a
HeLa cell cDNA library revealed that one of these 5' UTR-interacting
factors turns out to be the cellular protein GRSF-1 (362).
In vitro and in vivo binding analyses demonstrated that GRSF-1 can
specifically bind to the NP 5' UTR but not select NP 5' UTR mutants or
cellular RNA 5'UTRs (362) (Fig.
12A). Importantly, recombinant GRSF-1
was found to specifically stimulate the translation of an NP 5'
UTR-driven template in cell-free translation systems. Furthermore, the
translation efficiency of NP 5' UTR-driven templates was markedly
reduced in GRSF-1-depleted HeLa cell extracts but was restored by
GRSF-1-in reconstituted extracts. Competition experiments using NP 5'
UTR sequences similarly demonstrated a requirement for GRSF-1 binding
in the translation of viral but not cellular mRNA (Fig. 12B). Taken
together, these results demonstrate a specific interaction between
GRSF-1 and the influenza virus 5' UTR. More importantly, these results
suggest that influenza virus may recruit GRSF-1 to the 5' UTR to ensure
the selective translation of viral mRNAs in infected cells.
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Temporal regulation of influenza virus mRNA translation. In addition to regulating cellular mRNA translation, influenza virus carries out temporal regulation of its own viral gene expression at the level of translation. For example Yamanaka et al. (486, 487) transfected HeLa cells with CAT reporter gene appended to the 5' UTR of each of the viral mRNAs separately and superinfected these cells with influenza virus. They measured CAT activity at early and late times after infection. When the CAT construct contained the 5' UTR of an mRNA encoding an early protein such as NS, CAT activity was increased early after infection. Conversely when this was done with a late viral protein such as neuraminidase, CAT activity was higher at late times after infection. This avenue of investigation has not been pursued further, and exact sequences or trans-acting factors responsible for this regulation have not been identified. More recently, however, it was found that the influenza virus NS1 protein could stimulate the synthesis of the viral M1 protein (102). Site-directed mutagenesis studies showed that specific sequences within the M1 mRNA 5' UTR are required for this stimulation. These data, taken together with results discussed above, suggest a key role for the 5' UTR of influenza virus mRNAs in dictating temporal and selective translation in the infected cell.
Maintenance of translation in influenza virus-infected
cells.
Not only does influenza virus have to exert preferential
translation of its viral mRNAs, but also it has to ensure that the cell remains optimally translationally competent during infection (244). Without both these major strategies, viral
replication might be compromised, a scenario unacceptable to an
actively replicating cytopathic virus. The translational competence of
the infected cell is assured because influenza virus has developed an
intricate strategy to repress the activity of the PKR protein kinase.
That influenza virus represses PKR was first demonstrated by analyzing cells doubly infected with influenza virus and the adenovirus VA
RNAI-negative mutant dl331 (246, 247). In cells
infected by dl331 alone, there was a dramatic decline in the
levels of both viral and cellular protein synthesis (458).
This was due to excessive phosphorylation of the
eIF2 subunit by an active PKR, which cannot be inhibited due to the
absence of the virus-encoded VA RNAI (reviewed by Mathews and Shenk
[323]). In contrast, when dl331-infected
cells were superinfected with influenza virus, a reduction of the
protein kinase activity normally detected during dl331
infection was observed (246, 247). These data provided the
first evidence that influenza virus encodes or activates a gene product
that, analogous to VA RNAI, inhibits PKR and prevents any resultant
inhibition of protein synthesis initiation. It was subsequently shown
that the suppression of PKR activity also occurs in cells infected by
influenza virus alone (253).
Recruitment of P58IPK and inhibition of eIF2
phosphorylation during influenza virus infection.
A number of eukaryotic viruses have devised one or more strategies
to minimize the deleterious effects on protein synthesis caused by
activation of PKR (for a recent review, see reference 131). Unlike the strategies used by other viruses
described above, influenza virus utilizes at least two strategies,
involving a cellular and a viral protein, to block PKR activity. During
viral infection, influenza virus mobilizes a cellular protein, termed P58IPK, to repress PKR activity by blocking both the
autophosphorylation of PKR and the subsequent
phosphorylation of eIF2 by an active form of PKR
(294, 295). In uninfected cells, P58IPK
appears to form an inactive complex with its own inhibitor,
termed I-P58IPK (296). In response to activating
stimuli, such as viral infection or other cellular stresses,
P58IPK is released from its inhibitor. As a result, PKR
activity is repressed by a physical interaction between
P58IPK and PKR (130, 135, 378). To further
complicate the story, two different inhibitors of P58IPK,
the molecular chaperone Hsp40 (332) and another cellular
protein P52IPK (128), have recently been
identified. It has been postulated that these independent
P58IPK complexes are regulated by distinct cellular
pathways (128). Once released from its inhibitor,
P58IPK may negatively regulate PKR activity through
multiple steps, including the recruitment of molecular chaperone Hsp70
to inhibit kinase activity (333) and disruption of PKR
dimerization (447).
Role of the influenza virus NS1 protein in viral mRNA translation. The NS1 protein of influenza virus is an RNA-binding factor that inhibits both the nuclear export of poly(A)-containing mRNA and the splicing of pre-mRNA (86, 119, 311, 385). In addition to binding to poly(A) mRNA and a stem-bulge region in U6 small nuclear RNA, the NS1 protein binds to dsRNA (183). Evidence has accumulated to indicate that NS1 is involved in the translation of select viral mRNAs, including those encoding the viral matrix and nucleocapsid proteins (361). In this regard, NS1 is thought to interact with the viral 5' UTR to selectively stimulate the initiation of viral mRNA translation (244). To shed some light on how such translational selectivity may occur, recent evidence now indicates that NS1 can also form a complex with eIF4G in extracts from influenza virus-infected cells (T. Aragon, S. de la Luna, I. Novoa, L. Carrasco, J. Ortin, and A. Nieto, submitted for publication). Thus, we may hypothesize that NS1 can recruit eIF4G to the viral 5' UTR to facilitate translation initiation. Such an interaction could also contribute to the host shutoff due to competition with cellular mRNAs for eIF4G. Clearly, the exact mechanisms of NS1 specificity in mediating selective viral mRNA translation remain to be determined. It will be interesting to examine whether NS1 may interact with GRSF-1 to synergistically promote selective viral mRNA translation.
NS1 may also function during influenza virus infection to block the actions of PKR. Given the RNA-binding properties of NS1 and its ability to bind dsRNA, it was hypothesized that NS1 could compete with PKR in influenza virus-infected cells for binding to dsRNA. This tenet is supported by the observation that NS1 inhibits the activation of PKR and, as a result, the phosphorylation of eIF2 in vitro (312).
Furthermore, the NS1 protein also blocks the inhibition of translation
caused by dsRNA-mediated activation of PKR in reticulocyte
lysate extracts. The relevant role of NS1 in PKR regulation is further
strengthened by the finding that the two proteins can form a specific
complex in vitro (448). An inactive mutant of NS1, which
lacks a functional RNA-binding domain, was unable to bind to PKR.
Moreover, a PKR mutant defective in dsRNA binding did not
interact with or inhibit the NS1 protein in vivo. These results suggest
that NS1 exerts its effect, at least in part, through
heterodimerization with PKR, possibly in an RNA-dependent manner.
However, discretion is advised when using dsRNA-binding
mutants of PKR to determine the mechanism of these interactions because
both the dsRNA-binding and protein interaction properties of
PKR are closely embedded in the same regions. At any rate, the fact
that NS1 plays an in vivo role in modulating PKR function was recently
demonstrated by studies of mutant influenza viruses with a defective
NS1 protein (184). These variant viruses could not block the
activation of PKR in infected cells, leading to enhanced
phosphorylation of eIF2 and suppression of mRNA
translation. Furthermore, the level of phosphorylation
of PKR and eIF2 was well correlated with the defect in virus protein
synthesis. Consistent with its role in translation modulation, the NS1
protein has been shown to stimulate the translation of viral mRNAs
(86, 102), although it has not been determined if the PKR
pathway is involved. Collectively, these results suggest that NS1 may
facilitate viral replication by blocking the critical PKR-dependent arm
of the cellular IFN response. This idea is supported by work by
Garcia-Sastre et al., who used reverse genetics to engineer a
recombinant influenza virus, termed delNS1, which lacks the NS1 gene
(139). Similar to wild-type influenza virus, delNS1
replicated to high titer within cells deficient in IFN signaling
pathways. However, delNS1 replication was severely limited in cells in
which IFN signaling remained intact. Similarly, delNS1 replicated to
lethal titers in mice with a target deletion in the STAT1
gene, which renders cells unable to respond to IFN (77,
334). IFN-competent control mice effectively suppressed delNS1
replication. Thus, the NS1 gene of influenza virus may not play a
direct role in viral replication, but, rather, it functions to block
the antiviral effects of the host IFN system. In this regard, NS1 may
confer translational competence to influenza virus by removing the
translational blockade imposed by PKR.
Influenza virus appears to encode more than one strategy to repress PKR
function (Table 4). Influenza virus is
known to generate large amounts of both negative-strand and
positive-strand viral RNAs during infection, forming dsRNAs
that are capable of activating PKR. It seems logical that the virus
uses a cellular factor (P58IPK) and a viral protein (NS1)
to inhibit the activation of PKR in order to ensure efficient
protection against the resulting inhibition of translation that would
block virus replication. Alternatively, some of these PKR inhibitors
may interfere with activities of PKR not directly related to the
regulation of protein synthesis. Perhaps such viral multistrategies are
designed to fine-tune the activities of the enzyme at different stages
in the viral life cycle. Furthermore, since PKR is found in both the
nucleus and cytoplasm (68) the use of NS1, a predominantly
nuclear protein (166), and P58IPK, a cytoplasmic
protein (268), may allow influenza virus to control PKR
functions in both cellular compartments. Several key questions remain
to be addressed in future studies. (i) How does influenza virus
activate the P58IPK-PKR regulatory pathway? (ii) Would
cells devoid of P58IPK be more susceptible to influenza
virus infection and replication? (iii) Do NS1 and P58IPK
work in a synergistic manner to inhibit PKR function and enhance the
translation of viral mRNAs?
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VIRAL MODIFICATION OF CELLULAR FACTORS
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Viral replication requires a large amount of energy and thereby demands almost total metabolic control of cellular resources. As discussed in the previous sections, the competition for resources imposed by these conditions has created a playing field in which viruses have evolved mechanisms to supersede cellular mRNA translation through the recruitment and/or modification of translation factor function. Many of these processes of translation factor modification may have arisen through the efforts by viruses to block cellular countermeasures aimed at disrupting viral mRNA translation. As described in this section, the effects of translation factor modification range from altering the efficiency of cap-dependent translation and translation elongation to altering the rate of global mRNA translation and the efficacy of the innate antiviral response of the host cell.
Inactivation of eIF4E and modulation of the eIF4E-binding proteins. eIF4E may be considered the pivotal translation initiation factor. Compared to the other translation factors, it is present in limiting amounts in the cell, where it is required for assembly of the 5' cap-binding complex. As described above, eF4E binds to the 5' ends of both cellular and viral mRNAs and interacts with eIF4A and eIF4G. This macromolecular complex constitutes eIF4F and facilitates the binding of the mRNA to the 43S preinitiation complex (335). The affinity of eIF4E for the mRNA 5' cap is increased by phosphorylation on serine 209 and occurs in response to mitogenic stimulation (237, 431, 432). Analyses of eIF4E activity in the presence or absence of serine 209 phosphorylation has indicated that phospho-eIF4E is stimulatory for mRNA translation (432). The cellular enzyme responsible for serine 209 phosphorylation has been putatively identified as the Mnk1 protein kinase. Mnk1 was shown to phosphorylate eIF4E on serine 209 in vitro (473). More recent studies demonstrated that overexpression of Mnk1 could induce high levels of eIF4E phosphorylation in vivo (474), although the effects of Mnk1 overexpression on mRNA translation were not directly examined. These results are consistent with previous observations demonstrating that mitogen-induced eIF4E phosphorylation was dependent on the activity of the extracellular signal-related kinase and mitogen-activated protein kinase (339, 341, 472). Thus, Mnk1 may link eIF4E phosphorylation and translation stimulation through the mitogen-activated protein kinase-signaling pathway (387). In general, eF4E function is a requisite for cap-dependent translation but is thought to play a more critical role in the translation of mRNAs that have long 5' UTRs with regions of extensive secondary structure that are nonconducive to ribosomal scanning. These mRNAs are translated inefficiently due to limitations in eIF4E and the associated helicase activity of the assembled eIF4F complex (reviewed by Sonenberg [432]). Serine 209 phosphorylation is thought to increase the translational efficiency of these mRNAs by stimulating eIF4E cap-binding activity and increasing the level of mRNA-bound eIF4F helicase activity sufficiently to melt mRNA secondary structure.
The activity of eIF4E is also regulated through direct interaction with the eIF4E-binding proteins, 4E-BP1 to 4E-BP3 (367, 380). The 4E-BPs are low-molecular-weight proteins that link cap-dependent translation to mitogenic signaling pathways and are themselves directly regulated by phosphorylation (105, 305, 367). A model for eIF4E regulation by the 4E-BPs has been presented (387, 432). This model is based on the observations that interaction of eIF4E with 4E-BP1 disrupted the assembly of an eIF4E-eIF4G complex (172). 4E-BPs may facilitate the suppression of cap-dependent translation by binding to eIF4E and preventing the formation of an active eIF4F complex. Phosphorylation of the 4E-BP results in dissociation of the eIF4E-4E-BP inhibitory complex (367), thereby stimulating mRNA translation (387). 4E-BP phosphorylation and translation stimulation occur in response to mitogenic signaling and other cell growth-modulatory stimuli (154, 469). Elucidation of such cellular signaling cascades is currently an intense area of research. Viral disruption of eIF4E regulation may therefore have far-reaching consequences beyond supporting viral replication, including implications for apoptosis and oncogenic transformation (432). Viruses have targeted the processes of eIF4E regulation to facilitate translational selectivity for viral mRNAs during infection. In mammalian cells, the level of eIF4E serine 209 phosphorylation is reduced during infection with a number of viruses, including adenovirus and influenza virus (110, 244, 411, 499). In short, the block in eIF4E phosphorylation corresponds to a decrease in the level of host mRNA translation with little or no effects on translation of viral mRNAs. The molecular mechanisms by which eIF4E phosphorylation is reduced by influenza virus are not clear. Examination of mRNA translation in cells infected with influenza virus revealed that (i) virus infection significantly reduced the extent of eIF4E phosphorylation and the pool of active eIF4E and (ii) the efficiency of viral mRNA translation was insensitive to the resulting low levels of functional eIF4E and the eIF4F complex (110). All influenza virus mRNAs contain a short, conserved 5' UTR predicted to possess limited, if any, secondary structure (274). This short linear 5' UTR directs viral mRNA translation with a remarkably high efficiency. Such "strong" mRNAs may be less sensitive to limitations in the availability of eIF4F and associated helicase activity imposed by eIF4E dephosphorylation. As a result, dephosphorylation of eIF4E during influenza virus infection may favor the translation of viral mRNAs over the host mRNA and therefore may contribute to the host shutoff phenomenon. At this point, it is not known if influenza virus induces an eIF4E phosphatase activity, inhibits an eIF4E kinase including possibly Mnk1, or simply makes eIF4E unavailable for phosphorylation. These possibilities should provide a fertile area for future research in viral translational control mechanisms. In cells infected with adenovirus, the block in eIF4E phosphorylation plays a dual role of contributing to the host shutoff during late-stage infection and selecting for the translation (by ribosome shunt) of viral mRNAs that contain the tripartite leader. The actual mechanism by which adenovirus prevents eIF4E phosphorylation is not clear. Evidence from experiments that examined infection kinetics and levels of eIF4E phosphorylation has implicated a late adenovirus gene function in the coordinate reduction in eIF4E phosphorylation (499). An attractive hypothesis is that adenovirus may directly or indirectly inhibit Mnk1 or another cellular protein kinase(s) that phosphorylates eIF4E. Viral regulation of eIF4E function also occurs at the level of 4E-BP activity. Both poliovirus and EMCV inhibit 4E-BP phosphorylation within infected cells (155). Inhibition of 4E-BP phosphorylation may therefore contribute to the host shutoff of protein synthesis observed during picornavirus infection. This idea is supported by analyses of 2A-pro deficient strains of EMCV. Loss of 2A-pro decreased the efficiency of viral protein synthesis and abolished virus-induced host shutoff in infected cells (441). Interestingly, mutant viruses exhibited enhanced viral replication and increased efficiency of viral mRNA translation during infection in the presence of rapamycin and wortmannin, chemical inhibitors of 4E-BP phosphorylation (25). Thus, inhibition of 4E-BP phosphorylation complements EMCV mutations in 2A-pro to rescue viral mRNA translation (441). Together, these results indicate that inhibition of 4E-BP and repression of eIF4E function contributes to the host shutoff induced by picornavirus infection (26).Cleavage of eIF4G. With the exception of HAV, the host shutoff induced during picornavirus infection is extremely intense and results in nearly complete disruption of cellular mRNA translation to favor IRES-mediated translation of the viral mRNA. A main feature of this host shutoff involves disruption of eIF4F function through virus-mediated cleavage of eIF4G (Fig. 5), although it is now clear that other factors, including inhibition of 4E-BP phosphorylation, play a role in the shutoff process (174, 468). Cleavage of eIF4G proceeds in part through the actions of the virus-encoded 2A-pro during EMCV and poliovirus infections or through the actions of protease-L in foot-and-mouth disease virus infection (reviewed in references 197, 258, and 340). Cleavage of eIF4G effectively selects for cap-independent, IRES-mediated viral mRNA translation by removing the competition for the translational machinery imposed by cap-dependent cellular mRNA translation.
The implications of eIF4G cleavage upon host cap-dependent mRNA translation are best understood by examining eIF4G structure and function. eIF4G has been cloned from several different species and is present as structurally distinct isoforms that contain binding sites for interaction with eIF4E, eIF4A, eIF3, PABP, and the Mnk1 protein kinase (153, 214, 258, 340). The current model for eIF4F function in cap-dependent translation proposes that eIF4G serves as a molecular bridge for the assembly of the cap-binding complex upon the 5' end of the mRNA and facilitates its interaction with the 43S preinitiation complex. Furthermore, eIF4G may potentiate eIF4E phosphorylation and increased eIF4E activity by recruiting Mnk1 to the eIF4F complex (153, 383, 474). Poliovirus 2A-Pro cleaves eIF4G to yield discrete amino- and carboxyl-terminal cleavage products (Fig. 14). This prevents assembly of a functional eIF4F complex, thereby disrupting an essential step in cap-dependent translation initiation. Interestingly, cap-independent viral protein synthesis of select picornaviruses is stimulated by the carboxyl-terminal eIF4G cleavage product, which contains the binding sites for eIF3 and eIF4A (340, 354).
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Cleavage of PABP: disruption of the closed-loop translation complex. In addition to cleavage of eIF4E, picornaviruses may disrupt host mRNA translation through the modulation or modification of PABP. PABP is a 70-kDa RNA-binding protein that has remained highly conserved in evolution and plays a direct role in mRNA stability through its interaction with the poly(A) tails of mRNA (225). Recent evidence indicates that PABP directly participates in mRNA translation by functioning to bring the 3' UTR of the mRNA into the proximity of the 5' cap and the cap-binding complex. As described above, this circularization of the translation initiation complex is facilitated through PABP interactions with the mRNA poly(A) tail and eIF4G. The resulting "closed-loop" translation initiation complex is thought to stabilize assembled initiation factors and increase translation efficiency (225, 401). Thus, viral disruption of PABP function can be expected to (i) alter mRNA stability and (ii) reduce the overall rate and efficiency of cap-dependent mRNA translation.
Interestingly, it now appears that PABP is targeted for cleavage by 2A-Pro of the enteroviruses, coxsackievirus, and poliovirus. Analysis of PABP during coxsackievirus infection revealed that it was proteolytically cleaved during infection (259). In these studies, PABP was efficiently cleaved in vitro and in vivo by the virus-encoded 2A-Pro. Cleavage of human PABP occurred in a unique position that resulted in separation of the RNA-binding activity from the homodimerization activity. Importantly, the proteolytic fragments were inefficient at stimulating mRNA translation, suggesting that PABP cleavage may impart host translational suppression. Similar observations were extended to poliovirus infection, in which PABP cleavage was shown to occur through the actions of the viral 2A-Pro and, to a lesser extent, the viral 3C protease (233). In both studies PABP cleavage was accompanied by a dramatic loss of cellular translational activity in vitro and in vivo (233, 259). Taken together, these results suggest that direct cleavage of PABP may contribute to the inhibition of host mRNA translation during enterovirus infection. However, the extent to which PABP cleavage contributes to host protein synthesis shutoff, versus cleavage of eIF4G, has not been directly compared. Moreover, using infected cells, it will now be important to separate the effects of PABP cleavage on mRNA stability from direct translation-regulatory effects due to disruption of the "closed-loop" translation complex. It is tempting to propose that the cleavages of the PABP and eIF4G isoforms occur at distinct stages during viral infection. In this case, PABP cleavage may function to disrupt ongoing host translation while eIF4G cleavage events may be directed toward blocking de novo cap-dependent translation during infection.Modification of EF-1.
In addition to modifying the
components necessary for translation initiation, viruses may target
the process of translation elongation to facilitate the efficient
translation of viral mRNA. Translation factor EF-1 catalyzes the
critical step of delivering the aminoacyl-tRNAs to the elongating
ribosome. The viral regulatory protein, ICP0, forms a stable complex
with EF-1 during HSV infection (256). Analysis of the
translation efficiency of a reporter protein in RRL revealed that
the addition of recombinant ICP0 repressed translation in a
dose-dependent fashion. Although the actual rates of translation
elongation and the efficiency of translation initiation were not
addressed, it was concluded that ICP0 may function during specific
stages of HSV infection to modulate the translation of cellular
and viral mRNAs. Interestingly, EF-1 exists as hypo- and
hyperphosphorylated isoforms within HSV-infected cells
(256). Analysis of HSV mutants suggests that EF-1
phosphorylation is mediated by a protein kinase
encoded by the product of the viral U(L)13 gene
(257). Although a physiological role for
U(L)13-mediated phosphorylation of eEF-1
during HSV infections has not been demonstrated, it is interesting to
speculate that EF-1 phosphorylation may promote
viral protein synthesis by inducing the coordinate dissociation of
EF-1 from ICP0 at a specific point(s) during viral replication. Finally, EF-1 has also been shown to interact with the Tat protein of HIV-1 (485). In this case, the Tat-EF-1 interaction
resulted in a dramatic reduction in the efficiency of cellular but not viral mRNA translation. As with HSV, however, confirmation of EF-1
regulation by Tat, and its physiological role during HIV infection
remain an open question.
Disruption of eIF2 phosphorylation.
Eukaryotic cells respond to stress conditions, including viral
infection, in part by down-modulating the overall rate of protein synthesis. This translational control response to stress occurs largely
through the modification of eIF2. eIF2 functions to deliver the
Met-tRNAi to the 40S ribosome, and this constitutes a
rate-limiting step to translation initiation when eIF2 is modified
through phosphorylation by specific cellular
serine-threonine protein kinases. Currently, at least five
distinct eukaryotic protein kinases have been identified that play a
role in translational control by modulating eIF2 function; these are
the HRI, PERK, PEK, and PKR enzymes and the GCN2 protein kinase
(423). Known as the eIF2 protein kinase family, these enzymes respond to specific signals to phosphorylate serine 51 of
eIF2. Functional analyses of the eIF2 protein kinases indicate that each enzyme provides the cell with a unique ability to modulate mRNA translation in response to specific cellular stresses
(reviewed in references 201 and
477). For example, the HRI protein kinase is
expressed in mammalian reticulocytes and mediates eIF2 phosphorylation in response to heme depletion
(58). PERK and PEK reside on the endoplasmic
reticulum, where they mediate translational control in
response to endoplasmic reticulum stress (178, 419).
The yeast GCN2 enzyme presents an example of both global and specific control of mRNA translation by phosphorylating eIF2 in response to amino acid starvation (202). This results in the specific stimulation of GCN4 translation and concomitant repression of global
protein synthesis (201). GCN4 stimulates amino acid
production by inducing the expression of amino acid-biosynthetic
components. PKR is ubiquitously expressed in most mammalian
tissues. As described below, PKR is a component of the IFN-induced
cellular antiviral response and a pleiotropic mediator of
extracellular signals (68). However, PKR is best known
for its ability to phosphorylate eIF2 and repress mRNA
translation. Phosphorylation of eIF2 on serine 51 by PKR or the
other eIF2 protein kinases inhibits the guanine nucleotide exchange
reaction on eIF2 (Fig. 15). The
resulting eIF2 [S51-phospho]-GDP binary complex has a higher affinity
for the eIF2B guanine nucleotide exchanger than does the
nonphosphorylated eIF2 isoform. The increased affinity for
eIF2B impedes eIF2B function and results in sequestration of eIF2B
within an inactive complex with eIF2 [S51-phospho]-GDP. eIF2B
sequestering blocks the requisite recycling of GDP for GTP on
eIF2 and prevents de novo eIF2-GTP-Met-tRNAi ternary-complex
formation. As a result, mRNA translation initiation is
blocked.
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(i) PKR structure and function.
PKR and its role in
cellular metabolism have been extensively reviewed (58, 66,
201, 335, 404). Similar to many protein kinases, PKR is regulated
through a combination of transcriptional and posttranscriptional
processes including regulatory interactions with P58IPK
(135, 294, 378). PKR, however, is unique among the protein kinase superfamily in that it is the target for regulation
by virus-encoded inhibitory molecules (241-244, 321,
322). Structurally, PKR is composed of an
NH2-terminal regulatory domain and a COOH-terminal protein
kinase catalytic domain (Fig. 16A)
(68, 165, 336, 459). Ubiquitously expressed at low levels in
virtually all mammalian tissues examined (15), PKR is
transcriptionally induced by IFNs, which are secreted by host tissues
in response to viral infection (69) (see below). PKR is
rapidly activated after binding dsRNA or even single-stranded
RNA species that possess regions of extensive secondary structure
(70, 165, 248, 254, 316, 325, 331). This clearly imposes
problems for many eukaryotic viruses, which possess dsRNA
within the virion or produce high levels of dsRNA intermediates during replication. PKR binds dsRNA via its two NH2-terminal regulatory domain-binding motifs (dsRBM)
(Fig. 16A) (113, 165, 224, 363, 395). As a result of
these events, the kinase may undergo a conformational alteration which
triggers the catalytic activities of PKR (see reference
68 for a detailed review of PKR structure and
function). Typically, low levels of dsRNA are required to
initiate PKR activation while high levels of dsRNA actually
result in inhibition of PKR activation and suppression of kinase
function (70). Once bound to activator dsRNA,
PKR autophosphorylates on several critical serine and threonine
residues (453), rendering the kinase active. Recent evidence
by Zhu et al. (503) indicates that in addition to mediating
dsRNA activation of PKR, the dsRBMs function to target PKR to
the ribosome, thereby potentially providing access to the PKR
substrate, eIF2 (335). Activation of PKR can also proceed
through interaction with Pact, a novel PKR-activating protein that also
binds dsRNA (366). However, Pact appears to
activate PKR by a process that is independent of dsRNA, which
presumably involves alteration of PKR conformation.
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, consistent
with its reported roles in growth factor and calcium-mediated signal
transduction (344, 435), the regulation of transcription (266, 277, 317, 488), and the induction of apoptosis
(87, 293, 490) (reviewed by Proud [382] and
Williams [481]). However, it is the
phosphorylation of eIF2 by PKR that is most
frequently targeted for regulation by viruses. A diagram of the events
leading to PKR activation and eIF2 phosphorylation
is shown in Fig. 16B. IFNs, produced in response to viral infection,
induce the transcriptional activation of PKR, resulting in a
high level of PKR expression. During virus infection, the binding of
virus-encoded dsRNA to PKR initiates the PKR activation
process, which includes the aforementioned dimerization and
autophosphorylation events (84, 301, 373). Once activated, PKR phosphorylates eIF2 to limit mRNA
translation. As a result of these PKR-mediated processes, viral
replication is blocked at the level of protein synthesis (Fig. 15 and
16). High levels of eIF2 phosphorylation lead to an
antiproliferative state (19, 61, 89, 337, 395). Such a
situation would be incompatible with viral infection and replication,
providing another reason why viruses must inhibit PKR.
(ii) Mechanisms of PKR inhibition by eukaryotic viruses.
To avoid the deleterious effects on viral replication due to
PKR-mediated eIF2 phosphorylation, many viruses have
developed successful strategies to block PKR function, thus avoiding,
at least in part, the antiviral effects of IFN. As summarized in Table
4, viruses have employed a range of mechanisms to inhibit PKR function,
from targeting the PKR activation process to regulation of catalytic
function and beyond. These include directing inhibitors that (i)
interfere with the dsRNA-mediated activation of PKR, usually
by binding to the conserved dsRNA-binding domains or
sequestering RNA activators; (ii) interfere with kinase dimerization;
(iii) block the kinase catalytic site and PKR-substrate interactions; and (iv) alter the physical levels of PKR; and (v) may regulate eIF2
phosphorylation directly, or affect components
downstream from eIF2. Each PKR-inhibitory group includes a diverse
set of viruses, largely unrelated except for their ability to inhibit the kinase. In addition, some viruses employ multiple strategies for
inhibiting PKR, resulting in pleiotropic effects on PKR function. It is
noteworthy that such a diverse range of viruses has utilized often-limited genomic resources to target PKR for regulation. This reflects the pivotal role played by the kinase within the IFN-induced antiviral response of the host cell.
Disruption of the IFN-induced cellular antiviral response through inhibition of PKR. Upon infecting the cell of a vertebrate host, viruses must overcome the innate antiviral response provided by the cellular IFN system. IFNs are a family of cytokines which are secreted by the cells of vertebrate animals in response to viral infection and other cellular stresses. Once exposed to IFN, responsive cells initiate a signaling cascade which culminates in the specific induction of multiple IFN-inducible genes (88), only a small subset of which have been extensively characterized (for a review of the IFN system, the reader is directed to references 230, 346, 374, 403, 414, 415, and 437). The IFN-induced gene products, which play a role in fighting virus infection, include the 2'-5' oligoadenylate synthetase (28, 422), RNase L (421, 500), the Mx proteins (275, 369), and PKR (15, 212, 336, 451, 459). As a result, IFNs direct a block in viral gene expression at multiple levels, including the inhibition of viral RNA transcription and translation, and the degradation of viral transcripts.
Paramount to the IFN-induced antiviral response is the function of PKR, which not only imposes a limitation on viral mRNA translation but also participates in dsRNA- and IFN-induced signaling events (Fig. 16). These functional properties of PKR greatly contribute to the ability of IFN to provide the body's first level of defense against viral infection. PKR is required for the induction of IRF-1 activation (261, 277), which in turn regulates the transcription of a variety of IFN-inducible genes (260), including the type 1 IFNs, IFN- and IFN-
(277, 415, 488). PKR also contributes to the IFN-mediated
response through its ability to activate the transcription factor,
NF-
B (277, 297). Not surprisingly, inhibition of PKR
function results in attenuation of the IFN response by blocking one or
more of these PKR-dependent events (277, 488). Thus, PKR
presents an attractive target for virus-mediated inhibition of the host
IFN response (Fig. 16).
(i) Viral inhibition of PKR: HCV. The implications stemming from viral inhibition of PKR and disruption of the IFN response are perhaps best demonstrated by very recent work on the clinically relevant HCV. HCV now infects more than 2% of the worldwide population, including over 4 million in the United States (6). HCV infection often assumes a persistent course, which can lead to chronic hepatitis and liver cirrhosis, and is strongly associated with the development of hepatocellular carcinoma and lymphoproliferative disorders (355, 356, 463).
HCV infection is currently treated by parenteral administration of type I IFN alone or in combination with ribiviran, a nucleoside analog (326). Problematically, an increasingly high proportion of HCV-infected individuals (60 to 80%) fail to respond to IFN therapy or relapse after therapy cessation (208, 213). Response to IFN therapy differs among the six HCV genotypes but is observed, at some level, in all HCV genotypes worldwide. In an effort to understand the molecular mechanism(s) of HCV-mediated resistance to IFN therapy, several research groups have focused attention on sequencing clinical isolates of HCV from individuals who did or did not respond to IFN therapy (103, 104, 240, 278). What is clear from these studies is that sequence variation from the prototypic IFN-resistant HCV J strain (240) within the nonstructural 5A (NS5A) protein of the HCV polyprotein cleavage product is associated with sensitivity to IFN in Japanese HCV 1B subtypes (103, 104, 278). Viral isolates with multiple amino acid substitutions within a region of NS5A, termed the IFN sensitivity-determining region (ISDR; amino acids 2209 to 2248), were eliminated from HCV-infected patients during IFN therapy, while those exhibiting the prototypic ISDR sequence were IFN resistant, persisting at therapy cessation. These results suggested that HCV NS5A may mediate viral sensitivity to IFN through specific sequences located within or around the ISDR. Accordingly, it was demonstrated that NS5A from IFN-resistant strains of HCV 1A and 1B can physically bind PKR by an ISDR-dependent mechanism to inhibit kinase function, implicating NS5A as a mediator of the IFN-resistant HCV phenotype (133). It was subsequently hypothesized that mutations within the ISDR may similarly disrupt NS5A function to render HCV sensitive to the PKR-mediated antiviral effects of IFN. Subsequent analyses revealed that ISDR sequence variants of NS5A corresponding to IFN-resistant and -sensitive clinical isolates of HCV 1B (103) exhibited differential abilities to control PKR function in vivo (129, 132, 134). These studies demonstrated that NS5A from IFN-resistant HCV disrupted a critical step of PKR activation, resulting in repression of PKR function. In contrast, clinically defined mutations within the ISDR abrogated the PKR-regulatory function of NS5A. Further experiments mapped the PKR-interactive domain of HCV NS5A to a 64-amino-acid motif that includes the ISDR and the immediate carboxyl-terminal flanking region (129). Thus, it now appears that HCV may mediate resistance to IFN in part by blocking the PKR-dependent arm of the IFN response, through a direct NS5A-PKR interaction. In support of this, mutations within the PKR-binding domain on NS5A, including those within the ISDR, may confer IFN sensitivity by disrupting the NS5A-PKR interaction. Recent work indicates this to be true; infection of stable cell lines expressing wt or mutant NS5A with VSV revealed striking differences in the sensitivity of VSV to IFN in the presence or absence of functional NS5A (134, 377). In these studies, VSV replicated to significantly high levels within IFN-treated cells expressing wt NS5A from IFN-resistant HCV. In contrast, VSV replication was suppressed in cell lines expressing a nonfunctional NS5A mutant, reflecting the innate sensitivity of VSV to the antiviral effect of IFN. Analyses of PKR function and eIF2
phosphorylation attributed the apparent rescue of VSV
replication to the NS5A-mediated inhibition of PKR and a
resulting block in eIF2 phosphorylation (134) (Fig. 17). Moreover,
very recent evidence indicates that HCV disruption of PKR function is
not restricted to effects on eIF2 phosphorylation
alone. NS5A expression also renders cells refractory to PKR-dependent
signaling (134; M. Gale, Jr., unpublished data).
Disruption of host PKR signaling pathways is now considered an
important mechanism by which HCV may facilitate its domination of the
host IFN response.
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ISDR) was assessed by single-dimension
isoelectric focusing of cell extracts and anti-eIF2
VIRAL PERSISTENCE AND TRANSLATIONAL CONTROL
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As is clear from the material presented above, a common theme in viral translation programming is the preferential selection for the translation of viral mRNAs at the expense of host mRNA translation. While such programming is often compatible with the life cycle of viruses that mediate rapid, lytic infection, such as influenza virus, poliovirus, and EMCV, it is not always compatible with viruses that mediate persistent infection. Little is known of the nature of viral translational programming as it pertains to persistent infection, although it clearly requires that host mRNA translation remain sufficient to sustain the host cell and support viral persistence. In this section, we will take a brief look at the relationship between translational programming and viral persistence. As described below, persistent infection may involve viral translational programming that impacts the host cell at several levels, including the control of translation, apoptosis, and cell growth-regulatory pathways.
Translational programming and maintenance of viral persistence. Persistent infection requires the virus to ensure that the host cell remains translationally competent. This is important not only for the synthesis of viral proteins but also for the synthesis of cellular proteins and continued cellular viability. Analyses of the mechanisms by which viruses may mediate persistence and latency suggest that host cell integrity and translational competence are maintained through (i) viral modulation of specific cellular mRNA translation and (ii) viral modification of host signaling and translational regulatory pathways. As described above, the continued synthesis of ribosomal proteins during HSV-1 infection can be considered critical for maintaining viral persistence and latency. Interestingly, the level of ribosomal protein S6 phosphorylation is modulated in response to HSV-1 infection, thereby favoring the translation of host 5' TOP RNAs, including those that encode the ribosomal proteins (164). Modification of S6 phosphorylation may thus facilitate translational competence, in part by ensuring that the synthesis of ribosomal proteins remain uncompromised during infection. The mechanisms by which HSV-1 induces the modification of S6 have not been determined. It is conceivable that viral infection results in activation of one or more cellular protein kinases that phosphorylate S6 and/or that a virus-encoded protein kinase may effect S6 phosphorylation.
In addition to maintaining host translational competence during persistent infection, viruses must ensure that the synthesis of their own proteins remains uncompromised. As presented above, many eukaryotic viruses encode mechanisms to repress the cellular PKR protein kinase and avoid the deleterious effect upon protein synthesis due to high levels of eIF2 phosphorylation
(131). Thus, inhibition of PKR-dependent eIF2
phosphorylation can be seen as a mechanism to ensure
overall translational competence during viral infection. Through the
subsequent targeting of specific translational processes, including the
cap-binding reaction, elongation, and termination, viruses can then
manipulate host mRNA translation to the overall benefit of viral
replication. However, the implications for viral disruption of
PKR-dependent eIF2 phosphorylation are far-reaching
and are not limited to the effects on mRNA translation. As
presented above, repression of PKR provides the added advantage of
avoiding, in large part, the antiviral effects of the host IFN
response. Moreover, viral inhibition of eIF2
phosphorylation may disrupt critical host apoptotic and
tumor suppressor pathways, which rely specifically on the ability to
modify eIF2 activity (67, 68, 450).
Translational control, persistent infection, and regulation
of host apoptosis.
What are the implications of blocking
PKR function and preventing the phosphorylation of
eIF2 during viral infection? Recent evidence suggests that
inhibition of PKR function is a key feature in the establishment of
persistent viral infection. Lau and colleagues (489)
examined EMCV infection in established cell lines that were deficient
in PKR expression or in control cells that expressed normal levels of
PKR. EMCV normally mediates a cytolytic infection in vitro that is
characterized by massive apoptosis of permissive cells.
However, EMCV infection of PKR-deficient cells conferred persistent
infection to this otherwise cytolytic virus. Although viral RNA
translation was not directly examined in these studies, the results
suggest that constitutive disruption of PKR-dependent eIF2
phosphorylation may facilitate the establishment of
persistent viral infection. Such a relationship between viral control
of PKR and establishment of persistent infection may prove important for HCV infection. HCV mediates persistent infection within a majority
of infected individuals (6). Persistent HCV infection is
strongly associated with the development of hepatocellular carcinoma
and lymphoproliferative disorders (355, 356, 463). Recent
observations now indicate that viral persistence and disruption of host
apoptosis are linked to the block in eIF2
phosphorylation mediated by the HCV NS5A protein
(134). Expression of NS5A in mammalian cells induces a block
in eIF2 phosphorylation and concomitant stimulation
of mRNA translation through NS5A-mediated repression of PKR. NS5A
may contribute to HCV persistence by removing the translational
blockade imposed by PKR-dependent eIF2
phosphorylation (Fig. 17). Consequently, however,
constitutive expression of NS5A rendered cells refractory to
PKR-dependent apoptosis (134). These results support
previous studies indicating that eIF2
phosphorylation is an important component of cellular
apoptotic signaling (436). The mechanisms by which eIF2
phosphorylation promote apoptosis are not well
understood. Current thinking proposes that the
phosphorylation of eIF2 is required to block the
synthesis of antiapoptotic gene products during the
apoptotic response (450). More recently, however,
high-level PKR expression and PKR-dependent eIF2
phosphorylation have been associated with increased
synthesis of proapoptotic effector proteins, including Bax
and Fas (16, 95). Thus, eIF2 phosphorylation may result in the selective translation
of specific proapoptotic mRNAs. Constitutive disruption
of eIF2 phosphorylation that may occur during
persistent HCV infection may therefore render the host cell refractory
to apoptotic signaling. It is problematic for the host that
disruption of eIF2 phosphorylation and the ensuing
block in apoptotic signaling is associated with oncogenic transformation (16, 134).
Cell growth control, eIF2
phosphorylation, and oncogenic transformation.
Results from recent studies suggest that eIF2
phosphorylation may regulate cell growth, in part by
enforcing a translational control and apoptosis checkpoint on
cell proliferation. By this model, oncogenic potential is conferred to
persistent viral infections in which the eIF2 checkpoint is targeted
and constitutively disrupted. Thus, it is interesting that a wide range
of tumorigenic viruses possess mechanisms to repress PKR-dependent
eIF2 phosphorylation during infection (reviewed in
reference 131). Recent work suggests that PKR
activity and eIF2 phosphorylation are strictly
regulated during the cell division cycle (495;
M. Gale, Jr., C. Zhou, E. J. Firpo, M. G. Katze, A. Rudendsky, and B. R. Franza, Jr., submitted for publication)
and that inhibition of PKR function results in perturbation of cell
cycle control (16, 95). Our studies suggest that the NS5A
protein from IFN-resistant HCV may confer an oncogenic potential to
infected cells through the constitutive inhibition of PKR
(134). It is fitting to speculate that inhibition of PKR function and disruption of eIF2 phosphorylation may
contribute to the development of hepatocellular carcinoma in patients
persistently infected with HCV.
MRNA TRANSLATION AS A TARGET FOR
ANTIVIRAL THERAPY
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As our understanding of viral mRNA translation increases, it is becoming quite clear that viruses often utilize rather unorthodox tactics to ensure their efficient mRNA translation. Such viral strategies that deviate from the processes of conventional cellular mRNA translation may represent potential targets for the therapeutic intervention in viral replication. The utility of targeting translation for the development of antimicrobial therapeutics has been successfully demonstrated through the use of antibiotics such as tetracycline and erythromycin, which target bacterium-specific elements of translation. The challenges to developing successful antiviral therapeutics that target viral translational programming remain in (i) identifying translational targets that are specific to the virus and (ii) ensuring that host translation remains selectively undisturbed by the therapeutic intervention. As is evident from the numerous examples cited in this review, viral translational programming intersects with nearly every aspect of cellular mRNA translation, thereby making these challenges all the more spectacular. However daunting a task, therapeutic intervention in viral mRNA translation remains a promising arena for the development of effective antiviral compounds, which in large part have been limited to viral polymerase or protease inhibitors.
Recent work has used antisense oligonucleotide strategies, dsRNA selection and targeting strategies, and direct mutagenesis of viral mRNA to demonstrate that effective viral translational targeting can be achieved. Here we briefly present some interesting highlights in this developing field. This section features strategies that are under development as potential therapeutics to combat the global HCV pandemic. The development of antiviral drugs for HCV infection is problematic, due to the high mutation rate of HCV and the enormous level of quasispeciation that occurs during infection (46, 92). Thus, one can expect that potential HCV therapeutics that target the viral polymerase, helicase, or protease activities may ultimately be of limited value, since drug-resistant strains will certainly emerge. Indeed, this has already been demonstrated for HIV, which exhibits a similarly high mutation rate during infection (92). The roles of the HCV 5' and 3' UTRs in viral mRNA translation and replication and the fact that these regions are highly conserved among the different HCV genotypes make them viable targets for the development of antiviral therapies. Thus, strategies which target the function of the HCV IRES and the translation-stimulatory activity of the viral 3' UTR may represent efficient means of blocking HCV protein synthesis. Moreover, the development of compounds that may disrupt the ability of the NS5A protein to bind and repress PKR may serve to increase the efficacy of IFN and the existing IFN therapeutic regimens. For further information on therapeutic targeting of viral mRNA translation, the reader is referred to a review by Harford, and references therein (182).
IRES-mediated translation is not a common feature among cellular mRNAs, suggesting that it may represent a viable target for therapeutic intervention in viral mRNA translation. In support of this idea, Dasgupta and colleagues have identified a cellular RNA that possess an IRES-inhibitory function (467). This 60-nt RNA, termed IRNA, was isolated from S. cerevisiae and was first identified by examining the efficiency of poliovirus IRES-mediated translation in S. cerevisiae. These investigators found that poliovirus IRES translation was blocked in yeast and that this was due to a trans-acting factor that could similarly prevent poliovirus IRES translation when added to translationally competent HeLa extracts (79). Purified IRNA was shown to specifically inhibit IRES-mediated translation without having any effect on cap-dependent translation of cellular mRNAs. Interestingly, the IRNA was found to bind the cellular La protein (80), a major IRES-binding protein and effector of IRES-mediated translation (23). Evidence for other IRNA-polypeptide interactions was also demonstrated. These results suggested that the IRNA functions as an IRES-binding protein competitor to block translation from the poliovirus IRES. Subsequent studies revealed that expression of the IRNA in hepatoma cell lines similarly rendered a block to translation from the HCV IRES (81). Moreover, cells expressing the IRNA were refractory to infection with chimeric poliovirus under control of the HCV IRES. These results support the idea that the poliovirus and HCV IRES elements may bind a similar repertoire of cellular proteins. Structure-function analysis of the IRNA indicates that specific secondary structures are required for IRNA to bind cellular factors that promote IRES function (467). Thus, it appears that the IRNA may functionally mimic the IRES, at least in the context of secondary structure, and thereby compete for cellular proteins that mediate IRES translation. However, many questions remain to be addressed regarding the nature and origin of the IRNA itself. What is the cellular function of this RNA species, and are there homologous sequences present in the cells of higher eukaryotes? How may this IRNA control or interfere with cellular gene expression? Until such questions are answered, the potential therapeutic value of IRNA sequences will be significantly limited.
The targeted disruption of HCV genome translation has similarly been achieved using antisense oligonucleotides. Evaluation of the translation-inhibitory properties of a limited library of chemically modified oligonucleotides, directed to various regions of the HCV IRES and core protein-coding region, identified at least two antisense sequences that effectively inhibited HCV gene expression (175). In these studies, the expression of a truncated HCV genome in immortalized human hepatocytes was ablated by hybridization of an antisense oligonucleotide corresponding to a region encompassing the initiator AUG codon of the HCV core protein. Analysis of HCV RNA levels revealed that inhibition of genome expression was achieved without reducing genomic RNA expression. These results indicate that inhibition of HCV genome expression occurred at the level of translation and was not dependent on the activation of endogenous RNase H activity by duplex RNA. The translational block imposed by antisense oligonucleotides has yet to be confirmed by polyribosome analyses of HCV genome expression in the presence or absence of oligonucleotide treatment.
The possible utility of antisense oligonucleotides as an HCV antiviral therapy was demonstrated in a mouse model of HCV infection (498). This system utilized a recombinant vaccinia virus expressing an HCV 5' UTR-core region construct fused to the firefly luciferase gene. Translation of this HCV-core-luciferase construct was under control of the authentic HCV IRES. Mice infected with this vaccinia virus recombinant exhibited a block in liver-specific luciferase activity when treated with antisense oligonucleotides directed to the core protein initiation codon and flanking sequences. Luciferase expression remained high in infected mice that received oligonucleotide controls. These results are subject to the following criticisms: (i) they used a heterologous virus system, which may not faithfully represent events of HCV infection, and (ii) the actual mechanism(s) contributing to inhibition of HCV (luciferase) expression was not determined. However, considering the enormous number of applications of antisense strategies and the efficiency with which antisense transcripts disrupt gene expression (2, 262), antisense targeting of HCV replication remains a viable means of developing anti-HCV therapies. Taken together, these results indicate that antisense oligonucleotides may provide a potent mechanism by which to target viral mRNA translation as an antiviral therapy.
Another possible target of antisense-oligonucleotide strategies may be found within the HCV 3' UTR. As detailed above, this region of the HCV genome is highly conserved in all viral isolates, where it is thought to play an important a role in genome replication. It is appropriate to speculate that antisense oligonucleotides directed to within the HCV 3' UTR may disrupt transcription and block the translation-stimulatory activity induced by PTB binding (218). Finally, it should be noted that the use of antisense-oligonucleotide strategy to target specific RNAs for degradation by the IFN-induced RNase, RNase L, has been achieved (317, 461). Although this strategy is not directly aimed at blocking viral mRNA translation, it presents a viable option for targeting the HCV RNA for specific degradation. In contrast to strategies that depend on RNA degradation by RNase H, a predominant nuclear protein, RNase L is a resident cytoplasmic enzyme and would be available to disrupt HCV replication, which also takes place in the cytoplasm (500). The possibilities for targeting the HCV 3' UTR and the use of the RNase L pathway remain exciting areas of research into antisense oligonucleotides.
Similar to the use of antisense oligonucleotides, ribozymes, which are enzymatic RNA molecules that catalyze the cleavage of RNA, can be constructed to target specific RNA sequences (145). Ribozymes have been shown to be effective in blocking translation directed from the HCV IRES, although this probably occurs indirectly by RNA cleavage. Ribozymes constructed to target conserved sites within the viral 5' UTR blocked the translation of a luciferase reporter protein under control of the HCV IRES and 5' UTR in a tissue culture system, with little or no apparent toxicity to the host cell (402). Ribozymes have the added advantage of being efficiently packaged and expressed by various viral vectors, including vaccinia virus, adeno-associated virus, and various retroviruses. Moreover, ribozymes have been effective in catalyzing the degradation of both positive and negative strands of the HCV RNA (478). Strand-specific targeting of HCV RNA by ribozymes may thus hold promise for blocking the emergence of drug-resistant stains of HCV by eliminating new RNA variants.
Viral resistance to the current IFN-based therapeutic regimes for HCV
infection is a major problem (121, 208, 263) and is in part
responsible for the high frequency of persistent infections within the
HCV-infected population. As described above, HCV resistance to IFN has
been attributed in part to viral repression of the IFN-induced protein
kinase PKR. The NS5A-PKR interaction may provide a useful therapeutic
target for increasing the sensitivity to IFN in individuals who
initially fail to respond to IFN therapy. Disruption of the NS5A-PKR
interaction may restore eIF2 phosphorylation and the
translational blockade imposed by PKR, thereby increasing the
antiviral activity of IFN. Moreover, it is conceivable that restoration of PKR-dependent eIF2 phosphorylation
may prove useful in reducing the incidence of cellular proliferative
disorders associated with persistent HCV infection, since our results
indicate that NS5A repression of PKR may provide an oncogenic potential to HCV (134).
CONCLUSIONS AND PERSPECTIVES
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Investigations into the mechanisms and controls of viral protein synthesis have led the way to understanding the processes of cellular mRNA translation. Analyses of viral systems have introduced us to a better understanding of cellular antiviral pathways and signaling processes that affect mRNA translation. It is now clear that the translational control is an intimate part of most, if not all, metabolic processes of the cell. With this said, it now becomes important to understand the mechanisms of translational specificity; that is, how do cells and viruses impart translational control of specific mRNAs in a sea of translationally competent transcripts, and how do the extracellular environment and environmental cues effect mRNA translation?
Selective mRNA translation is a hallmark of many viral infections. While it is clear that viruses often encode mechanisms to disrupt cellular mRNA translation, the molecular mechanisms of selective viral mRNA translation under conditions of host protein synthesis shutoff remain poorly understood. In particular, the molecular mechanisms of IRES-mediated translation have yet to be elucidated. What are the basal factors that are required to support IRES-mediated translation? What is the actual role of IRES-binding proteins, and how do they function to stimulate viral mRNA translation? Similar questions extend to the mechanisms of cap-dependent selective translation. Identification of trans-acting factors that directly interact with viral RNA and understanding the implications of such interactions for viral translational programming remain the logical course of action. However, these approaches have been limited in scope by the nature of the traditional biochemical and molecular techniques often used when conducting such studies.
Recent advances in protein and nucleic acid analyses, such as combined mass spectrometry and sequence database searching (170), tandem mass spectrometry (171), and the current genomic technologies (44, 56, 236, 299, 407), make these techniques viable tools for investigating the mechanisms of translation control. The application of such techniques should allow for characterizing novel RNA-protein interaction. Genomic technologies, such as the use of high-density genome arrays, have already offered spectacular and sometimes surprising insights into the differential spectrum of cellular gene expression under various environmental conditions (88, 308, 350, 375, 408, 434). Similar applications to assess the array of both viral and cellular gene expression in virus-infected cells are proving equally interesting. In particular, high-density genome arrays can be used for the parallel identification of transcriptionally and translationally regulated mRNAs (150, 502). Recent studies have demonstrated the utility of this latter approach. Morris and colleagues (504) separated cellular mRNAs from resting or mitogenically activated fibroblast cultures into discrete pools based on the number of mRNA-bound ribosomes. By interrogating cDNA microarray filters with probes generated from the mRNA pools, these investigators found that translational control of cellular mRNA, at least in the context of mitogenic stimulation, was remarkably selective and represented less than 1% of the mRNAs in this cross-sectional analysis. Taking this application a step further, Sarnow and colleagues (234) interrogated cDNA microarrays with probes derived from polyribosome-associated mRNAs prepared from poliovirus-infected cells. This application allowed the investigators to identify cellular mRNAs that could be translated independent of a functional eIF4F complex. Remarkably, it was found that approximately 2 to 3% of the mRNAs analyzed were associated with polyribosomes under these conditions. When examined for their ability to direct the cap-independent translation of a bicistronic reporter protein, it was shown that at least a subset of these mRNAs contained functional IRES sequences within their 5' UTR. An emerging theme from these analyses is that mRNAs, whose gene products have been implicated in a variety of stress responses, are translated with little or no requirement for eIF4F. Thus, IRES-mediated translation may be prevalent among mRNAs that are involved in acute cellular responses.
Development of an understanding of the acute cellular signaling pathways that modulate mRNA translation has now attracted the attention of those in the signal transduction field (43). It has become very clear that cellular mRNA translation is controlled in response to specific environmental signals that modulate development, cell proliferation, and apoptosis. Recent evidence now indicates that viruses impinge on these pathways during infection to promote viral replication. An important question now is how viral infection affects these pathways and how this may contribute to viral pathogenesis.
Determination of the three-dimensional structure of translation-regulatory proteins and translation factors represents another fruitful area for future research. Structural determination of translation factors is essential for a full understanding of the complexity and control of translation factor interactions and function. This was demonstrated by the recent elucidation of the crystal structure of eIF4E (318). As with eIF4E, translation factors play prominent roles in cell proliferation and malignancy (67, 267, 433). Understanding translation factor structure may pave the way for rational drug design of anticancer and antiviral therapeutic compounds.
ACKNOWLEDGMENTS
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We thank Marlene Wambach, Cecelia Boyer, and Dagma Daniel for their excellent administrative and technical support for the past several years. We thank the many members of the Katze laboratory, past and present, for their contributions to our work. We are grateful to individuals who have collaborated with us over the years and who continue to share our interests in translational control. We thank Young Woo Park for sharing major results prior to publication.
M.G. thanks the Helen Hay Whitney Foundation for outstanding postdoctoral support. Work in the Gale laboratory is funded by the UT Southwestern Endowed Scholars Program and by the Texas Applied Research Program. Work in the Katze laboratory is supported by National Institutes of Health grants AI22646, RR00166, and AI41629; by Ribogene Corporation; and by the Gustave and Louise Pfeiffer Research Foundation.
FOOTNOTES
* Corresponding author. Mailing address: Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9048. Phone: (214) 648-5940. Fax: (214) 648-5905. E-mail: mgale{at}mednet.swmed.edu.
REFERENCES
|
TopPrevious |
|---|
| 1. | Agol, V. I. 1998. Virus robustness and perseverance. Mol. Biol. 32:44-49. |
| 2. | Alama, A., F. Barbieri, M. Cagnoli, and G. Schettini. 1997. Antisense oligonucleotides as therapeutic agents. Pharmacol. Res. 36:171-178[CrossRef][Medline]. |
| 3. | Ali, N., and A. Siddiqui. 1995. Interaction of polypyrimidine tract-binding protein with the 5' noncoding region of the hepatitis C virus RNA genome and its functional requirement in internal initiation of translation. J. Virol. 69:6367-6375[Abstract]. |
| 4. |
Ali, N.,
C. Wang, and A. Siddiqui.
1995.
Translation of hepatitis C virus genome.
Princess Takamatsu Symp.
25:99-110 |
| 5. |
Alonso-Caplen, F. V.,
M. G. Katze, and R. M. Krug.
1988.
Efficient transcription, not translation, is dependent on adenovirus tripartite leader sequences at late times of infection.
J. Virol.
62:1606-1616 |
| 6. | Alter, M. J. H. 1997. Epidemiology of hepatitis C. Hepatology 26:62-65[CrossRef]. |
| 7. | Amoroso, P., M. Rapicetta, M. E. Tosti, A. Mele, E. Spada, S. Bounocore, G. Lettieri, P. Peirri, P. Chionne, A. R. Ciccaglione, and L. Sagliocca. 1998. Correlation between virus genotype and chronicity rate in acute hepatitis C. J. Hepatol. 28:939-944[CrossRef][Medline]. |
| 8. |
Andreansky, S.,
L. Soroceanu,
E. R. Flotte,
J. Chou,
J. M. Markert,
G. Y. Gillespie,
B. Roizman, and R. J. Whitley.
1997.
Evaluation of genetically engineered herpes simplex viruses as oncolytic agents for human malignant brain tumors.
Cancer Res.
57:1502-1509 |
| 9. | Reference deleted. |
| 10. | Aranda, M., and A. Maule. 1998. Virus-induced host gene shutoff in animals and plants. Virology 243:261-267[CrossRef][Medline]. |
| 11. |
Arrick, B. A.,
A. L. Lee,
R. L. Grendell, and R. Derynck.
1991.
Inhibition of translation of transforming growth factor-3 mRNA by its 5' untranslated region.
Mol. Cell. Biol.
11:4306-4313 |
| 12. | Atkins, J. F., and R. F. Gesteland. 1996. Regulatory recoding, p. 653-684. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 13. |
Avni, D.,
Y. Biberman, and O. Meyuhas.
1997.
The 5' terminal oligopyrimidine tract confers translational control on TOP mRNAs in a cell type- and sequence context-dependent manner.
Nucleic Acids Res.
25:995-1001 |
| 14. |
Babich, A.,
L. Feldman,
J. Nevins,
J. E. Darnell, and C. Weinberger.
1983.
Effect of adenovirus on metabolism of specific host mRNAs: transport control and specific translation discrimination.
Mol. Cell. Biol.
3:1212-1221 |
| 15. |
Baier, L. J.,
T. Shors,
S. T. Shors, and B. L. Jacobs.
1993.
The mouse antiphosphotyrosine immunoreactive kinase, TIK, is indistinguishable from the double-stranded RNA-dependent, interferon-induced protein kinase, PKR.
Nucleic Acids Res.
21:4830-4835 |
| 16. | Balachandran, S., C. N. Kim, W.-C. Yeh, T. W. Mak, and G. N. Barber. 1998. Activation of the dsRNA-dependent protein kinase, PKR, induces apoptosis through FADD-mediated death signaling. EMBO J. 17:6888-6902[CrossRef][Medline]. |
| 17. |
Banerjee, A. K.
1980.
5'-terminal cap structure in eucaryotic messenger ribonucleic acids.
Microbiol. Rev.
44:175-205 |
| 18. | Barber, G. N., M. Wambach, S. Thompson, R. Jagus, and M. G. Katze. 1995. Mutants of the RNA-dependent protein kinase (PKR) lacking double-stranded RNA binding domain I can act as transdominant inhibitors and induce malignant transformation. Mol. Cell. Biol. 15:3138-3146[Abstract]. |
| 19. |
Barber, G. N.,
M. Wambach,
M.-L. Wong,
T. E. Dever,
A. G. Hinnebusch, and M. G. Katze.
1993.
Translational regulation by the interferon-induced double-stranded-RNA-activated 68-kDa protein kinase.
Proc. Natl. Acad. Sci. USA
90:4621-4625 |
| 20. | Beattie, E., K. L. Denzler, J. Tartaglia, M. E. Perkus, E. Paoletti, and B. L. Jacobs. 1995. Reversal of the interferon-sensitive phenotype of a vaccinia virus lacking E3L by expression of the reovirus S4 gene. J. Virol. 69:499-505[Abstract]. |
| 21. |
Beattie, E.,
J. Tartaglia, and E. Paoletti.
1991.
Vaccinia virus-encoded eIF-2 homolog abrogates the antiviral effect of interferon.
Virology
183:419-422[CrossRef][Medline].
|
| 22. | Beloso, A., C. Martinez, J. Valcarcel, J. F. Santaren, and J. Ortin. 1992. Degradation of cellular mRNA during influenza virus infection: its possible role in protein synthesis shutoff. J. Gen. Virol. 73:575-581[Abstract]. |
| 23. | Belsham, G. J., and N. Sonenberg. 1996. RNA-protein interactions in regulation of picornavirus RNA translation. Microbiol. Rev. 60:499-511[Abstract]. |
| 24. | Benkirane, M., C. Neuveut, R. F. Chun, S. M. Smith, C. E. Samuel, A. Gatignol, and K.-T. Jeang. 1997. Oncogenic potential of TAR RNA binding protein TRBP and its regulatory interaction with RNA-dependent protein kinase PKR. EMBO J. 16:611-624[CrossRef][Medline]. |
| 25. | Beretta, L., A.-C. Gingras, Y. V. Svitkin, M. N. Hall, and N. Sonenberg. 1996. Rapamycin blocks the phosphorylation of 4E-BP1 and inhibits cap-dependent initiation of translation. EMBO J. 15:658-664[Medline]. |
| 26. | Beretta, L., Y. V. Svitkin, and N. Sonenberg. 1996. Rapamycin stimulates viral protein synthesis and augments the shutoff of host protein synthesis upon picornavirus infection. J. Virol. 70:8993-8996[Abstract]. |
| 27. |
Bernstein, P.,
S. W. Peltz, and J. Ross.
1989.
The poly(A)-poly(A)-binding protein complex is a major determinant of mRNA stability in vitro.
Mol. Cell. Biol.
9:659-670 |
| 28. | Bisbal, C., T. Salehzada, B. Lebleu, and B. Bayard. 1989. Characterization of two murine (2'-5')(A)n-dependent endonucleases of different molecular mass. Eur. J. Biochem. 179:595-602[Abstract]. |
| 29. |
Black, T.,
B. Safer,
A. G. Hovanessian, and M. G. Katze.
1989.
The cellular 68,000 Mr protein kinase is highly autophosphorylated and activated yet significantly degraded during poliovirus infection: implications for translational regulation.
J. Virol.
63:2244-2252 |
| 30. |
Black, T. L.,
G. N. Barber, and M. G. Katze.
1993.
Degradation of the interferon-induced 68,000-Mr protein kinase by poliovirus requires RNA.
J. Virol.
67:791-800 |
| 31. | Blight, K. J., A. A. Kolykhalov, K. E. Reed, E. V. Vagapov, and C. M. Rice. 1998. Molecular virology of hepatitis C virus: an update with respect to potential antiviral targets. Antiviral Ther. 3(Suppl. 3):71-81. [Medline] |
| 32. | Blight, K. J., and C. M. Rice. 1997. Secondary structure determination of the conserved 98-base sequence at the 3' terminus of hepatitis C virus genome RNA. J. Virol. 71:7345-7352[Abstract]. |
| 33. | Blondel, B., G. Duncan, T. Couderc, F. Delpeyroux, and F. ColbereGarapin. 1998. Molecular aspects of poliovirus biology with a special focus on the interactions with nerve cells. J. Neurovirol. 4:1-29[Medline]. |
| 34. |
Blyn, L. B.,
K. M. Swiderek,
O. Richards,
D. C. Stahl,
B. L. Semler, and E. Ehrenfeld.
1996.
Poly(rC) binding protein 2 binds to stem-loop IV of the poliovirus RNA 5' noncoding region: identification by automated liquid chromatography-tandem mass spectrometry.
Proc. Nat. Acad. Sci. USA
93:11115-11120 |
| 35. | Blyn, L. B., J. S. Towner, B. L. Semler, and E. Ehrenfeld. 1997. Requirement of poly(rC) binding protein 2 for translation of poliovirus RNA. J. Virol. 71:6243-6246[Abstract]. |
| 36. | Borman, A., M. T. Howell, J. G. Patton, and R. Jackson. 1993. The involvement of a spliceosome component in internal initiation of rhinovirus RNA translation. J. Gen. Virol. 74:1775-1788[Abstract]. |
| 37. | Borman, A. M., F. G. Deliat, and K. M. Kean. 1994. Sequences within the poliovirus internal ribosome entry segment control viral RNA synthesis. EMBO J. 13:3149-3157[Medline]. |
| 38. | Borovjagin, A., T. Pestova, and I. Shatsky. 1994. Pyrimidine tract binding protein strongly stimulates in vitro encephalomyocarditis virus RNA translation at the level of preinitiation complex formation. FEBS Lett. 351:299-302[CrossRef][Medline]. |
| 39. |
Brand, S. R.,
R. Kobayashi, and M. B. Mathews.
1997.
The Tat protein of human immunodeficiency virus type 1 is a substrate and inhibitor of the interferon-induced, virally activated protein kinase, PKR.
J. Biol. Chem.
272:8388-8395 |
| 40. | Brierley, I., M. R. Meredith, A. J. Bloys, and T. G. Hagervall. 1997. Expression of a coronavirus ribosomal frameshift signal in Escherichia coli: influence of tRNA anticodon modification on frameshifting. J. Mol. Biol. 270:360-373[CrossRef][Medline]. |
| 41. | Brown, B. A., and E. Ehrenfeld. 1979. Translation of poliovirus RNA in vitro: changes in cleavage pattern and initiation sites by ribosomal salt wash. Virology 97:396-405[CrossRef][Medline]. |
| 42. | Brown, E. A., S. P. Day, R. W. Jansen, and S. M. Lemon. 1999. The 5' nontranslated region of hepatitis A virus RNA: secondary structure and elements required for translation in vitro. J. Virol. 65:5828-5838. |
| 43. | Brown, E. J., and S. L. Schreiber. 1996. A signaling pathway to translational control. Cell 86:517-520[CrossRef][Medline]. |
| 44. | Brown, P. O., and D. Botstein. 1999. Exploring the new world of the genome with DNA microarrays. Nat. Genet. 21:33-37[CrossRef][Medline]. |
| 45. | Bruno, S., E. Silini, A. Crosignani, F. Borzio, G. Leandro, F. Bono, M. Asti, S. Rossi, A. Larghi, A. Cerino, M. Podda, and M. U. Mondelli. 1997. Hepatitis C virus genotypes and risk of hepatocellular carcinoma in cirrhosis: a prospective study. Hepatology 25:754-758[CrossRef][Medline]. |
| 46. | Bukh, J., R. Miller, and R. Purcell. 1995. Genetic heterogeneity of hepatitis C virus: quasispecies and genotypes. Semin. Liver Dis. 15:41-63[Medline]. |
| 47. | Buratti, E., M. Gerotto, P. Pontisso, A. Alberti, S. G. Tisminetzky, and F. E. Baralle. 1997. In vivo translational efficiency of different hepatitis C virus 5'-UTRs. FEBS Lett. 411:275-280[CrossRef][Medline]. |
| 48. | Cao, J., and A. P. Geballe. 1994. Mutational analysis of the translational signal in the human cytomegalovirus gpUL4 (gp48) transcript leader by retroviral infection. Virology 205:151-160[CrossRef][Medline]. |
| 49. | Cao, J., and A. P. Geballe. 1996. Inhibition of nascent-peptide release at translation termination. Mol. Cell. Biol. 16:7109-7114[Abstract]. |
| 50. | Cao, J., and A. P. Geballe. 1998. Ribosomal release without peptidyl tRNA hydrolysis at translation termination in a eukaryotic system. RNA 4:181-188[Abstract]. |
| 51. |
Carpick, B. W.,
V. Graziano,
D. Schneider,
R. K. Maitra,
X. Lee, and B. R. G. Williams.
1997.
Characterization of the solution structure between the interferon-induced double-stranded RNA-activated protein kinase and HIV-1 transactivating region RNA.
J. Biol. Chem.
272:9510-9516 |
| 52. |
Carroll, K.,
O. Elroy-Stein,
R. Moss, and R. Jagus.
1993.
Recombinant vaccinia virus K3L gene product prevents activation of double-stranded RNA-dependent, initiation factor 2-specific protein kinase.
J. Biol. Chem.
268:12837-12842 |
| 53. |
Cassady, K. A.,
M. Gross, and B. Roizman.
1998.
The second-site mutation in the herpes simplex virus recombinants lacking the gamma(1)34.5 genes precludes shutoff of protein synthesis by blocking the phosphorylation of eIF-2 alpha.
J. Virol.
72:7005-7011 |
| 54. |
Centrella, M., and J. Lucas-Lenard.
1982.
Regulation of protein synthesis in vesicular stomatitis virus-infected mouse L-929 cells by decreased protein synthesis initiation factor 2 activity.
J. Virol.
41:781-791 |
| 55. |
Chang, H.-W.,
J. C. Watson, and B. L. Jacobs.
1992.
The E3L gene of vaccinia virus encodes an inhibitor of the interferon-induced, double-stranded RNA-dependent protein kinase.
Proc. Natl. Acad. Sci. USA
89:4825-4829 |
| 56. |
Chee, M.,
R. Yang,
E. Hubbell,
A. Berno,
X. C. Huang,
D. Stern,
J. Winkler,
D. J. Lockhart,
M. S. Morris, and S. P. A. Fodor.
1996.
Accessing genetic information with high-density DNA arrays.
Science
274:610-614 |
| 57. |
Chen, C. Y., and P. Sarnow.
1995.
Initiation of protein synthesis by the eukaryotic translation apparatus on circular RNAs.
Science
268:415-417 |
| 58. |
Chen, J.-J., and I. M. London.
1995.
Regulation of protein synthesis by heme-regulated eIF-2 kinase.
Trends Biochem. Sci.
20:105-108[CrossRef][Medline].
|
| 59. |
Chen, X. Y.,
H. S. Kang,
L. X. Shen,
M. Chamorro,
H. E. Varmus, and I. Tinoco.
1996.
A characteristic bent conformation of RNA pseudoknots promotes 1 frameshifting during translation of retroviral RNA.
J. Mol. Biol.
260:479-483[CrossRef][Medline].
|
| 60. |
Choi, S. K.,
W. C. Merrick,
W. L. Zoll, and T. E. Dever.
1998.
Promotion of Met-tRNA(i)(Met) binding to ribosomes by yIF2, a bacterial IF2 homolog in yeast.
Science
280:1757-1760 |
| 61. | Chong, K. L., L. Feng, K. Schappert, E. Meurs, T. F. Donahue, J. D. Friesen, A. G. Hovanessian, and B. R. G. Williams. 1992. Human P68 kinase exhibits growth suppression in yeast and homology to the translational regulator GCN2. EMBO J. 11:1553-1562[Medline]. |
| 62. |
Chou, J.,
J. J. Chen,
M. Gross, and B. Roizman.
1995.
Association of a Mr 90,000 phosphoprotein with protein kinase PKR in cells exhibiting enhanced phosphorylation of translation initiation factor eIF2 and premature shutoff of protein synthesis after infection with 34.5 mutants of herpes simplex virus 1.
Proc. Natl. Acad. Sci. USA
92:10516-10520 |
| 63. |
Chung, S.,
M. Eckrich,
B. N. Perrone,
D. T. Kohn, and H. Furneaux.
1997.
The Elav-like proteins bind to a conserved regulatory element in the 3'-untranslated region of GAP-43 mRNA.
J. Biol. Chem.
272:6593-6598 |
| 64. | Clemens, M. J. 1993. The small RNAs of Epstein-Barr virus. Mol. Biol. Rep. 17:81-92[CrossRef][Medline]. |
| 65. | Clemens, M. J. 1994. Regulation of eukaryotic protein synthesis by protein kinases that phosphorylate initiation factor eIF-2. Mol. Biol. Rep. 19:201-210[CrossRef][Medline]. |
| 66. | Clemens, M. J. 1996. Protein kinases that phosphorylate eIF-2 and eIF-2B, and their role in eukaryotic cell translational control, p. 139-172. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 67. | Clemens, M. J., and U.-A. Bommer. 1999. Translational control: the cancer connection. Int. J. Biochem. Cell Biol. 31:1-23[CrossRef][Medline]. |
| 68. | Clemens, M. J., and A. Elia. 1997. The double-stranded RNA-dependent protein kinase PKR: structure and function. J. Interferon Cytokine Res. 17:503-524[Medline]. |
| 69. | Clemens, M. J., J. W. B. Hershey, A. G. Hovanessian, B. L. Jacobs, M. G. Katze, R. J. Kaufman, P. Lengyel, C. E. Samuel, G. C. Sen, and B. R. G. Williams. 1993. PKR: proposed nomenclature for the RNA-dependent protein kinase induced by interferon. J. Interferon Res. 13:241[Medline]. |
| 70. |
Clemens, M. J.,
K. G. Laing,
I. W. Jeffrey,
A. Schofield,
T. V. Sharp,
A. Elia,
V. Matys,
M. C. James, and V. J. Tilleray.
1994.
Regulation of the interferon-inducible eIF-2 protein kinase by small RNAs.
Biochimie
76:770-778[CrossRef][Medline].
|
| 71. | Collier, A. J., S. X. Tang, and R. M. Elliott. 1998. Translation efficiencies of the 5' untranslated region from representatives of the six major genotypes of hepatitis C virus using a novel bicistronic reporter assay system. J. Gen. Virology 79:2359-2366[Abstract]. |
| 72. | Corbin, A., and J. L. Darlix. 1996. Functions of the 5' leader of murine leukemia virus genomic RNA in virion structure, viral replication and pathogenesis, and MLV-derived vectors. Biochimie 78:632-638[CrossRef][Medline]. |
| 73. |
Cosentino, G. P.,
S. Venkatesan,
F. C. Serluca,
S. R. Green,
M. B. Mathews, and N. Sonenberg.
1995.
Double-stranded-RNA-dependent protein kinase and TAR RNA-binding protein form homo- and heterodimers in vivo.
Proc. Natl. Acad. Sci. USA
92:9445-9449 |
| 74. |
Craig, A. W.,
G. P. Cosentino,
O. Donzé, and N. Sonenberg.
1996.
The kinase insert domain of interferon-induced protein kinase PKR is required for activity but not for interaction with the pseudosubstrate K3L.
J. Biol. Chem.
271:24526-24533 |
| 75. | Craig, A. W., A. Haghighat, A. T. Yu, and N. Sonenberg. 1998. Interaction of polyadenylate-binding protein with the eIF4G homologue PAIP enhances translation. Nature 392:520-523[CrossRef][Medline]. |
| 76. | Cullen, B. R. 1991. Regulation of HIV-1 gene expression. FASEB J. 5:2361-2368[Abstract]. |
| 77. | Darnell, J. E., Jr. 1998. Studies of IFN-induced transcriptional activation uncover the Jak-Stat pathway. J. Interferon Cytokine Res. 18:549-554[Medline]. |
| 78. | Das, A. T., B. Klaver, B. F. Klasens, J. B. vanWamel, and B. Berkhout. 1997. A conserved hairpin motif in the R-U5 region of the human immunodeficiency virus type 1 RNA genome is essential for replication. J. Virol. 71:2346-2356[Abstract]. |
| 79. |
Das, S.,
P. Coward, and A. Dasgupta.
1994.
A small yeast RNA selectively inhibits internal initiation of translation programmed by poliovirus RNA: specific interaction with cellular proteins that bind to the viral 5'-untranslated region.
J. Virol.
68:7200-7211 |
| 80. | Das, S., D. J. Kenan, D. Bocskai, J. D. Keene, and A. Dasgupta. 1996. Sequences within a small yeast RNA required for inhibition of internal initiation of translation: interaction with La and other cellular proteins influences its inhibitory activity. J. Virol. 70:1624-1632[Abstract]. |
| 81. |
Das, S.,
M. Ott,
A. Yamane,
W. M. Tsai,
M. Gromeier,
F. Lahser,
S. Gupta, and A. Dasgupta.
1998.
A small yeast RNA blocks hepatitis C virus internal ribosome entry site (HCV IRES)-mediated translation and inhibits replication of a chimeric poliovirus under translational control of the HCV IRES element.
J. Virol.
72:5638-5647 |
| 82. |
Davies, M. V.,
H.-W. Chang,
B. L. Jacobs, and R. J. Kaufman.
1993.
The E3L and K3L vaccinia virus gene products stimulate translation through inhibition of the double-stranded RNA-dependent protein kinase by different mechanisms.
J. Virol.
67:1688-1692 |
| 83. |
Davies, M. V.,
O. Elroy-Stein,
R. Jagus,
B. Moss, and R. J. Kaufman.
1992.
The vaccinia virus K3L gene product potentiates translation by inhibiting double-stranded-RNA-activated protein kinase and phosphorylation of the alpha subunit of eukaryotic initiation factor 2.
J. Virol.
66:1943-1950 |
| 84. |
Davis, S., and J. C. Watson.
1996.
In vitro activation of the interferon-induced, double-stranded RNA-dependent protein kinase PKR by RNA from the 3' untranslated regions of human alpha-tropomyosin.
Proc. Natl. Acad. Sci. USA
93:508-513 |
| 85. |
Degnin, C. R.,
M. R. Schleiss,
J. Cao, and A. P. Geballe.
1993.
Translational inhibition mediated by a short upstream open reading frame in the human cytomegalovirus gpUL4 (gp48) transcript.
J. Virol.
67:5514-5521 |
| 86. | de la Luna, S., P. Fortes, A. Beloso, and J. Ortin. 1995. Influenza virus NS1 protein enhances the rate of translation initiation of viral mRNAs. J. Virol. 68:2427-2433. |
| 87. |
Der, S. D.,
Y.-L. Yang,
C. Weissman, and B. R. G. Williams.
1997.
A double-stranded RNA-activated protein kinase-dependent pathway mediating stress-induced apoptosis.
Proc. Natl. Acad. Sci. USA
94:3279-3283 |
| 88. |
Der, S. D.,
A. Zhou,
B. R. G. Williams, and R. H. Silverman.
1998.
Identification of genes differentially regulated by interferon alpha, beta, or gamma using oligonucleotide arrays.
Proc. Natl. Acad. Sci. USA
95:15623-15628 |
| 89. |
Dever, T. E.,
J.-J. Chen,
G. N. Barber,
A. M. Cigan,
L. Feng,
T. F. Donahue,
I. M. London,
M. G. Katze, and A. G. Hinnebusch.
1993.
Mammalian eukaryotic initiation factor eIF2 kinases functionally substitute for GCN2 protein kinase in the GCN4 translational control mechanism of yeast.
Proc. Natl. Acad. Sci. USA
90:4616-4620 |
| 90. | Diaz, J. J., D. Simonin, T. Masse, P. Deviller, K. Kindbeiter, L. Denoroy, and J. J. Madjar. 1993. The herpes simplex virus type 1 US11 gene product is a phosphorylated protein found to be non-specifically associated with both ribosomal subunits. J. Gen. Virol. 74:397-406[Abstract]. |
| 91. |
Dolph, P. J.,
J. Juang, and R. J. Schneider.
1990.
Translation by the adenovirus tripartite leader: elements which determine independence from cap-binding protein complex.
J. Virol.
64:2669-2677 |
| 92. | Domingo, E., E. Baranowski, C. M. Ruiz-Jarabo, A. M. Martin-Hernandez, J. C. Saiz, and C. Escarmis. 1998. Quasispecies structure and persistence of RNA viruses. Emerging Infect. Dis. 4:521-527[Medline]. |
| 93. | Reference deleted. |
| 94. |
Dominguez, D. I.,
L. A. Ryabova,
M. M. Pooggin,
W. Schmidt-Puchta,
J. Fütterer, and T. Hohn.
1998.
Ribosome shunting in cauliflower mosaic virus: identification of an essential and sufficient structural element.
J. Biol. Chem.
273:3669-3678 |
| 95. | Donze, O., J. Dostie, and N. Sonenberg. 1999. Regulatable expression of the interferon-induced double-stranded RNA dependent protein kinase PKR induces apoptosis and Fas receptor expression. Virology 256:322-329[CrossRef][Medline]. |
| 96. | Donze, O., R. Jagus, A. E. Koromilas, J. W. B. Hershey, and N. Sonenberg. 1995. Abrogation of translation initiation factor eIF-2 phosphorylation causes malignant transformation of NIH 3T3 cells. EMBO J. 14:3828-3834[Medline]. |
| 97. | Doohan, J. P., and C. E. Samuel. 1992. Biosynthesis of reovirus-specified polypeptides: ribosome pausing during the translation of reovirus S1 mRNA. Virology 186:409-425[CrossRef][Medline]. |
| 98. |
Doohan, J. P., and C. E. Samuel.
1993.
Biosynthesis of reovirus-specified polypeptides.
J. Biol. Chem.
268:18313-18320 |
| 99. |
Dorner, A. J.,
B. L. Semler,
R. J. Jackson,
R. Hanecak,
E. Duprey, and E. Wimmer.
1984.
In vitro translation of poliovirus RNA: utilization of internal initiation sites in reticulocyte lysate.
J. Virol.
50:507-514 |
| 100. | Ehrenfeld, E. 1982. Poliovirus-induced inhibition of host-cell protein synthesis. Cell 28:435-436[CrossRef][Medline]. |
| 101. | Ehrenfeld, E. 1996. Initiation of translation by picornavirus RNAs, p. 549-574. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 102. |
Enami, K.,
T. A. Sato,
S. Nakada, and M. Enami.
1994.
Influenza virus NS1 protein stimulates translation of the M1 protein.
J. Virol.
68:1432-1437 |
| 103. |
Enomoto, N.,
I. Sakuma,
Y. Asahina,
M. Kurosaki,
T. Murakami,
C. Yamamoto,
Y. Ogura,
N. Izumi,
F. Maruno, and C. Sato.
1996.
Mutations in the nonstructural protein 5A gene and response to interferon in patients with chronic hepatitis C virus 1b infection.
N. Engl. J. Med.
334:77-81 |
| 104. | Enomoto, N., I. Sakuma, Y. Asahina, M. Kurosaki, T. Murankami, C. Yamamoto, N. Izumi, F. Marumo, and C. Sato. 1995. Comparison of full-length sequences of interferon-sensitive and resistant hepatitis C virus 1b. J. Clin. Investig. 96:224-230[Medline]. |
| 105. |
Fadden, P.,
T. A. J. Haystead, and J. C. Lawrence.
1997.
Identification of phosphorylation sites in the translational regulator, PHAS-I, that are controlled by insulin and rapamycin in rat adipocytes.
J. Biol. Chem.
272:10240-10247 |
| 106. |
Fajardo, J. E., and A. J. Shatkin.
1990.
Translation of bicistronic viral mRNA in transfected cells: regulation at the level of elongation.
Proc. Natl. Acad. Sci. USA
87:328-332 |
| 107. |
Falcone, D., and D. W. Andrews.
1991.
Both the 5' untranslated region and the sequences surrounding the start site contribute to efficient initiation of translation in vitro.
Mol. Cell. Biol.
11:2656-2664 |
| 108. | Farabaugh, P. J. 1996. Programmed translational frameshifting. Annu. Rev. Genetics 30:507-528[CrossRef][Medline]. |
| 109. | Fehrmann, F., R. Welker, and H. G. Krausslich. 1997. Intracisternal A-type particles express their proteinase in a separate reading frame by translational frameshifting, similar to D-type retroviruses. Virology 235:352-359[CrossRef][Medline]. |
| 110. |
Feigenblum, D., and R. J. Schneider.
1993.
Modification of eukaryotic initiation factor 4F during infection by influenza virus.
J. Virol.
67:3027-3035 |
| 111. |
Felsenstein, K. M., and S. P. Goff.
1988.
Expression of the gag-pol fusion protein of Moloney murine leukemia virus without gag protein does not induce virion formation or proteolytic processing.
J. Virol.
62:2179-2182 |
| 112. |
Felsenstein, K. M., and S. P. Goff.
1992.
Mutational analysis of the gag-pol junction of Moloney murine leukemia virus: requirements for expression of the gag-pol fusion protein.
J. Virol.
66:6601-6611 |
| 113. |
Feng, G.-S.,
K. L. Chong,
A. Kumara, and B. R. G. Williams.
1992.
Identification of double-stranded RNA-binding domains in the interferon-induced double-stranded RNA-activated p68 kinase.
Proc. Natl. Acad. Sci. USA
89:5447-5451 |
| 114. | Fenwick, M. L., and J. Clark. 1982. Early and delayed shut-off of host protein synthesis in cells infected with herpes simplex virus. J. Gen. Virol. 61:121-125[Abstract]. |
| 115. | Fernadez, I., G. Castellano, M. J. Domingo, A. Fuertes, F. Colina, F. Canga, F. J. de la Cruz, A. G. de la Camara, and J. A. Solis. 1997. Influence of viral genotype and the level of viremia on the severity of liver injury and the response to interferon therapy. Scand. J. Gastroenterol. 32:7-76. |
| 116. | Fesq, H., M. Bacher, M. Nain, and D. Gemsa. 1994. Programmed cell death (apoptosis) in human monocytes infected by influenza virus. Immunobiology 190:175-182[Medline]. |
| 117. | Fields, B. N. 1990. Virology. Raven Press, New York, N.Y. |
| 118. |
Finco, T. S.,
J. K. Westwick,
J. L. Norris,
A. A. Beg,
C. J. Der, and A. S. Baldwin, Jr.
1997.
Oncogenic Ha-Ras-induced signaling activates NF-B transcriptional activity, which is required for cellular transformation.
J. Biol. Chem.
272:24113-24116 |
| 119. | Fortes, P., A. Beloso, and J. Ort'in. 1994. Influenza virus NS1 protein inhibits pre-mRNA splicing and blocks mRNA nucleocytoplasmic transport. EMBO J. 13:704-712[Medline]. |
| 120. | Frederickson, R. M., and N. Sonenberg. 1992. Signal transduction and regulation of translation initiation. Semin. Cell Biol. 3:107-115[Medline]. |
| 121. | Fried, M., and J. Hoofnagle. 1995. Therapy of hepatitis C. Semin. Liver Dis. 15:82-91[Medline]. |
| 122. |
Furuya, T., and M. M. C. Lai.
1993.
Three different cellular proteins bind to complementary sites on the 5'-end-positive and 3'-negative strands of mouse hepatitis virus RNA.
J. Virol.
67:7215-7222 |
| 123. |
Fütterer, J.,
K. Gordon,
J.-M. Bonneville,
H. Sanfacon,
B. Pisan,
J. R. Penswick, and T. Hohn.
1988.
Differential inhibition of downstream gene expression by the cauliflower mosaic virus 35S RNA leader.
Nucleic Acids Res.
16:8377-8390 |
| 124. | Fütterer, J., Z. Kiss-L'aszl'o, and T. Hohn. 1993. Nonlinear ribosome migration on cauliflower mosaic virus 35S RNA. Cell 73:789-802[CrossRef][Medline]. |
| 125. | Galabru, J., M. G. Katze, N. Robert, and A. G. Hovanessian. 1989. The binding of double-stranded RNA and adenovirus VAI RNA to the interferon-induced protein kinase. Eur. J. Biochem. 178:581-589[Abstract]. |
| 126. | Reference deleted. |
| 127. | Reference deleted. |
| 128. |
Gale, M., Jr.,
C. M. Blakely,
D. A. Hopkins,
M. W. Melville,
M. Wambach,
P. R. Romano, and M. G. Katze.
1998.
Regulation of interferon-induced protein kinase PKR: modulation of P58IPK inhibitory function by a novel protein, P52rIPK.
Mol. Cell. Biol.
18:859-871 |
| 129. |
Gale, M., Jr.,
C. M. Blakely,
B. Kwieciszewski,
S.-L. Tan,
M. Dossett,
M. J. Korth,
S. J. Polyak,
D. R. Gretch, and M. G. Katze.
1998.
Control of PKR protein kinase by hepatitis C virus nonstructural 5A protein: molecular mechanisms of kinase regulation.
Mol. Cell. Biol.
18:5208-5218 |
| 130. | Gale, M., Jr., and M. G. Katze. 1997. What happens inside lentivirus or influenza virus infected cells: insights into regulation of cellular and viral protein synthesis. Methods Companion Methods Enzymol. 11:383-401. [CrossRef] |
| 131. | Gale, M., Jr., and M. G. Katze. 1998. Molecular mechanisms of interferon resistance mediated by viral-directed inhibition of PKR, the interferon-induced protein kinase. Pharmacol. Ther. 78:29-46[CrossRef][Medline]. |
| 132. | Gale, M., Jr., M. J. Korth, and M. G. Katze. 1998. Repression of the PKR protein kinase by hepatitis C virus: a potential mechanism for interferon resistance. Clin. Diagn. Virol. 10:157-162[CrossRef][Medline]. |
| 133. | Gale, M., Jr., M. J. Korth, N. M. Tang, S.-L. Tan, D. A. Hopkins, T. E. Dever, S. J. Polyak, D. R. Gretch, and M. G. Katze. 1997. Evidence that hepatitis C virus resistance to interferon is mediated through repression of the PKR protein kinase by the nonstructural 5A protein. Virology 230:217-227[CrossRef][Medline]. |
| 134. |
Gale, M., Jr.,
B. Kwieciszewski,
M. Dossett,
H. Nakao, and M. G. Katze.
1999.
Anti-apoptotic and oncogenic potentials of hepatitis C virus are linked to interferon resistance by viral repression of the PKR protein kinase.
J. Virol.
73:6506-6516 |
| 135. | Gale, M., Jr., S.-L. Tan, M. Wambach, and M. G. Katze. 1996. Interaction of the interferon-induced PKR protein kinase with inhibitory proteins P58IPK and vaccinia virus K3L is mediated by unique domains: implications for kinase regulation. Mol. Cell. Biol. 16:4172-4181[Abstract]. |
| 136. | Gallie, D. R. 1991. The cap and poly(A) tail function synergistically to regulate mRNA translational efficiency. Genes Dev. 5:2108-2116[Abstract]. |
| 137. | Gallie, D. R. 1996. Translational control of cellular and viral mRNAs. Plant Mol. Biol. 32:145-158[CrossRef][Medline]. |
| 138. | Gallie, D. R. 1998. A tale of two termini: a functional interaction between the termini of an mRNA is a prerequisite for efficient translation initiation. Gene 216:1-11[CrossRef][Medline]. |
| 139. | Garcia-Sastre, A., A. Egorov, D. Matassov, S. Brandt, D. E. Levy, J. E. Durbin, P. Palese, and T. Muster. 1999. Influenza A virus lacking the NS1 gene replicates in interferon-deficient systems. Virology 252:324-330[CrossRef]. |
| 140. |
Garfinkel, M. S., and M. G. Katze.
1992.
Translational control by influenza virus: selective and cap-dependent translation of viral mRNAs in infected cells.
J. Biol. Chem.
267:9383-9390 |
| 141. | Garfinkel, M. S., and M. G. Katze. 1993. How does influenza virus regulate gene expression at the level of mRNA translation? Let us count the ways. Gene Expression 3:109-118[Medline]. |
| 142. |
Garfinkel, M. S., and M. G. Katze.
1993.
Translational control by influenza virus: selective translation is mediated by sequences within the viral mRNA 5'-untranslated region.
J. Biol. Chem.
268:22223-22226 |
| 143. | Garfinkel, M. S., and M. G. Katze. 1994. Influenza virus control of protein synthesis. Sci. Am. 1994(issue on Science and Medicine):64-73. |
| 144. | Garry, R. F., J. M. Bishop, S. Parker, K. Westbrook, G. Lewis, and M. R. F. Waite. 1979. Na+ and K+ concentrations and the regulation of protein synthesis in sindbis virus-infected chick cells. Virology 96:108-120[CrossRef][Medline]. |
| 145. | Gaughan, D. J., and A. S. Whitehead. 1999. Function and biological applications of catalytic nucleic acids. Biochim. Biophys. Acta 1445:1-20[Medline]. |
| 146. | Geballe, A. P. 1996. Translational control mediated by upstream AUG codons, p. 173-197. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory, Plainview, N.Y. |
| 147. |
Geballe, A. P., and E. S. Mocarski.
1988.
Translational control of cytomegalovirus gene expression is mediated by upstream AUG codons.
J. Virol.
62:3334-3340 |
| 148. | Geballe, A. P., and D. R. Morris. 1994. Initiation codons within 5'-leaders of mRNAs as regulators of translation. Trends Biochem. Sci. 19:159-164[CrossRef][Medline]. |
| 149. | Gehrke, L., L. E. Hann, and R. L. Kaspar. 1994. Translational control of gene expression. Annu. Rep. Med. Chem. 29:248-254. |
| 150. | Geiss, G. K., R. E. Bumgarner, M. An, M. B. Agy, A. van't Wout, E. Hammersmark, V. Carter, D. Upchurch, J. I. Mullins, and M. G. Katze. 2000. Large-scale monitoring of host cell gene expression during HIV-1 infection using cDNA microarrays. Virology 266:8-16[CrossRef][Medline]. |
| 151. | Gesteland, R. F., and J. F. Atkins. 1996. Recoding: dynamic reprogramming of translation. Annu. Rev. Biochem. 65:741-768[CrossRef][Medline]. |
| 152. |
Giege, R.,
M. Sissler, and C. Florentz.
1998.
Universal rules and idiosyncratic features in tRNA identity.
Nucleic Acids Res.
26:5017-5035 |
| 153. | Gingras, A.-C., B. Raught, and N. Sonenberg. 1999. eIF4 initiation factors: effectors of mRNA recruitment to ribosomes and regulators of translation. Annu. Rev. Biochem. 68:963. |
| 154. |
Gingras, A. C.,
S. G. Kennedy,
M. A. OLeary,
N. Sonenberg, and N. Hay.
1998.
4E-BP1, a repressor of mRNA translation, is phosphorylated and inactivated by the Akt(PKB) signaling pathway.
Genes Dev.
12:502-513 |
| 155. |
Gingras, A. C.,
Y. Svitkin,
G. J. Belsham,
A. Pause, and N. Sonenberg.
1996.
Activation of the translational suppressor 4E-BP1 following infection with encephalomyocarditis virus and poliovirus.
Proc. Natl. Acad. Sci. USA
93:5578-5583 |
| 156. | Glass, M. J., and D. F. Summers. 1992. A cis-acting element within the hepatitis A virus 5' non-coding region required for in vitro translation. Virus Res. 26:15-31[CrossRef][Medline]. |
| 157. | Godefroy-Colburn, T., C. Thivent, and L. Pink. 1985. Translational discrimination between the four RNAs of alfalfa mosaic virus: a quantitative evaluation. Eur. J. Biochem. 147:541-548[Abstract]. |
| 158. | Goldenberg, D., N. Mador, M. J. Ball, A. Panet, and I. Steiner. 1997. The abundant latency-associated transcripts of herpes simplex virus type 1 are bound to polyribosomes in cultured neuronal cells and during latent infection in mouse trigeminal ganglia. J. Virol. 71:2897-2904[Abstract]. |
| 159. | Goldenberg, D., N. Mador, A. Panet, and I. Steiner. 1998. Tissue specific distribution of the herpes simplex virus type 1 latency-associated transcripts on polyribosomes during latent infection. J. Neurovirol. 4:426-432[Medline]. |
| 160. | Gottlieb, E., and J. A. Stelitz. 1989. Function of the mammalian La protein: evidence for its action in transcription termination by RNA polymerase III. EMBO J. 8:851-861[Medline]. |
| 161. |
Gradi, A.,
Y. V. Svitkin,
H. Imataka, and N. Sonenberg.
1998.
Proteolysis of human eukaryotic translation initiation factor eIF4GII, but not eIF4GI, coincides with the shutoff of host protein synthesis after poliovirus infection Proc.
Natl. Acad. Sci. USA
95:11089-11094 |
| 162. |
Graff, J.,
J. Cha,
L. B. Blyn, and E. Ehrenfeld.
1998.
Interaction of poly(rC) binding protein 2 with the 5' noncoding region of hepatitis A virus RNA and its effects on translation.
J. Virol.
72:9668-9675 |
| 163. | Gray, N. K., and M. Wickens. 1998. Control of translation initiation in animals. Annu. Rev. Cell Dev. Biol. 14:399-458[CrossRef][Medline]. |
| 164. | Greco, A., A. M. Laurent, and J. J. Madjar. 1997. Repression of beta-actin synthesis and persistence of ribosomal protein synthesis after infection of HeLa cells by herpes simplex virus type 1 infection are under translational control. Mol. Gen. Genet. 256:320-327[CrossRef][Medline]. |
| 165. | Green, S. R., and M. B. Mathews. 1992. Two RNA binding motifs in the double-stranded RNA activated protein kinase, DAI. Genes Dev. 6:2478-2490[Abstract]. |
| 166. |
Greenspan, D.,
P. Palese, and M. Krystal.
1988.
Two nuclear location signals in the influenza virus NS1 nonstructural protein.
J. Virol.
62:3020-3026 |
| 167. |
Gromeier, M.,
L. Alexander, and E. Wimmer.
1999.
Internal ribosome entry site substitution eliminates meurovirulence in intergeneric poliovirus recombinants.
Proc. Natl. Acad. Sci. USA
93:2370-2375 |
| 168. |
Gromeier, M.,
B. Bossert,
M. Arita,
A. Nomoto, and E. Wimmer.
1999.
Dual stem-loops within the poliovirus internal ribosomal entry site control neurovirulence.
J. Virol.
73:958-964 |
| 169. |
Gunnery, S. A.,
P. Rice,
H. D. Robertson, and M. B. Mathews.
1990.
Tat-responsive region RNA of human immunodeficiency virus 1 can prevent activation of the double-stranded RNA-activated protein kinase.
Proc. Natl. Acad. Sci. USA
87:8687-8691 |
| 170. | Gygi, S. P., D. K. M. Han, A.-C. Gingras, N. Sonenberg, and R. Aebersold. 1999. Protein analysis by mass spectrometry and sequence database searching: tools for cancer research in the post-genomic era. Electrophoresis 20:310-319[CrossRef][Medline]. |
| 171. |
Gygi, S. P.,
Y. Rochon,
B. R. Franza, and R. Aebersold.
1999.
Correlation between protein and mRNA abundance in yeast.
Mol. Cell. Biol.
19:1720-1730 |
| 172. | Haghighat, A., S. Mader, A. Pause, and N. Sonenberg. 1995. Repression of cap-dependent translation by 4E-binding protein 1: competition with p220 for binding to eukaryotic initiation factor-4E. EMBO J. 14:5701-5709[Medline]. |
| 173. |
Haghighat, A., and N. Sonenberg.
1997.
eIF4G dramatically enhances the binding of eIF4E to the mRNA 5'-cap structure.
J. Biol. Chem.
272:21677-21680 |
| 174. | Haghighat, A., Y. Svitkin, I. Novoa, E. Kuechler, T. Skern, and N. Sonenberg. 1996. The eIF4G-eIF4E complex is the target for direct cleavage by the rhinovirus 2A proteinase. J. Virol. 70:8444-8450[Abstract]. |
| 175. | Hanecak, R., V. BrownDriver, M. C. Fox, R. F. Azad, S. Furusako, C. Nozaki, C. Ford, H. Sasmor, and K. P. Anderson. 1996. Antisense oligonucleotide inhibition of hepatitis C virus gene expression in transformed hepatocytes. J. Virol. 70:5203-5212[Abstract]. |
| 176. |
Hanks, S. K.,
A. M. Quinn, and T. Hunter.
1988.
The protein kinase family: conserved features and deduced phylogeny of the catalytic domains.
Science
241:42-52 |
| 177. | Hann, L. E., A. C. Webb, J. M. Cai, and L. Gehrke. 1997. Identification of a competitive translation determinant in the 3' untranslated region of alfalfa mosaic virus coat protein mRNA. Mol. Cell. Biol. 17:2005-2013[Abstract]. |
| 178. | Harding, H. P., Y. Zhang, and D. Ron. 1999. Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature 397:271-274[CrossRef][Medline]. |
| 179. | Hardwick, J. M. 1997. Virus-induced apoptosis. Adv. Pharmacol. 41:295-336[Medline]. |
| 180. |
Hardwicke, M. A., and R. M. Sandri-Goldin.
1994.
The herpes simplex virus regulatory protein ICP27 contributes to the decrease in cellular mRNA levels during infection.
J. Virol.
68:4797-4810 |
| 181. |
Hardy, W. R., and R. M. Sandri-Goldin.
1994.
Herpes simplex virus inhibits host cell splicing, and regulatory protein ICP27 is required for this effect.
J. Virol.
68:7790-7799 |
| 182. | Harford, J. B. 1995. Translation-targeted therapeutics for viral diseases. Gene Expression 4:357-367[Medline]. |
| 183. | Hatada, E., and R. Fukuda. 1992. Binding of influenza A virus NS1 protein to dsRNA in vitro. J. Gen. Virol. 73:3325-3329[Abstract]. |
| 184. |
Hatada, E.,
S. Saito, and R. Fukuda.
1999.
Mutant influenza viruses with a defective NS1 protein cannot block the activation of PKR in infected cells.
J. Virol.
73:2425-2433 |
| 185. | Hatfield, D., Y. X. Feng, B. J. Lee, A. Rein, G. J. Levin, and S. Oroszlan. 1989. Chromatographic analysis of the aminoacyl-tRNAs which are required for translation of codons at and around the ribosomalframeshift sites of HIV, HTLV-1, and BLV. Virology 173:736-742[CrossRef][Medline]. |
| 186. | Hatfield, D., and S. Oroszlan. 1990. The where, what and how of ribosomal frameshifting in retroviral protein synthesis. Trends Biochem. Sci. 15:186-190[CrossRef][Medline]. |
| 187. |
Hayes, B. W.,
G. C. Telling,
M. M. Myat,
J. F. Williams, and S. F. Flint.
1990.
The adenovirus L4 100-kDa protein is necessary for efficient translation of viral late mRNA species.
J. Virol.
64:2732-2742 |
| 188. |
He, B.,
J. Chou,
R. Brandimarti,
I. Mohr,
Y. Gluzman, and B. Roizman.
1997.
Suppression of the phenotype of 134.5 herpes simplex virus 1: failure of activated RNA-dependent protein kinase to shut off protein synthesis is associated with a deletion in the domain of the 47 gene.
J. Virol.
71:6049-6054[Abstract].
|
| 189. | He, B., J. Chou, D. A. Liebermann, B. Hoffman, and B. Roizman. 1996. The carboxyl terminus of the murine MyD116 gene substitutes for the corresponding domain of the gamma(1)34.5 gene of herpes simplex virus to preclude the premature shutoff of total protein synthesis in infected human cells. J. Virol. 70:84-90[Abstract]. |
| 190. |
He, B.,
M. Gross, and B. Roizman.
1997.
The gamma(1)34.5 protein of herpes simplex virus I complexes with protein phosphatase 1 alpha to dephosphorylate the alpha subunit of the eukaryotic translation initiation factor 2 and preclude the shutoff of protein synthesis by double-stranded RNA-activated protein kinase.
Proc. Natl. Acad. Sci. USA
94:843-848 |
| 191. |
Hellen, C. U. T.,
T. V. Pestova,
M. Litterst, and E. Wimmer.
1994.
The cellular polypeptide p57 (pyrimidine tract-binding protein) binds to multiple sites in the poliovirus 5' nontranslated region.
J. Virol.
68:941-950 |
| 192. | Hellen, C. U. T., and E. Wimmer. 1994. Translation of encephalomyocarditis virus RNA: parameters influencing the selection of the internal initiation site. EMBO J. 13:1673-1681[Medline]. |
| 193. | Hellen, C. U. T., and E. Wimmer. 1995. Translation of encephalomyocarditis virus RNA by internal ribosome entry. Curr. Top. Microbiol. Immunol. 203:31-63[Medline]. |
| 194. |
Hellen, C. U. T.,
H. W. Witherall,
M. Schmid,
S. H. Shin,
T. V. Pestova,
A. Gil, and E. Wimmer.
1993.
A cytomplasmic 57-kDa protein that is required for translation of picornavirus RNA by internal ribosome entry is identical to the nuclear pyrimidine tract-binding protein.
Proc. Natl. Acad. Sci. USA
90:7642-7646 |
| 195. | Hemmings-Mieszczak, M., G. Steger, and T. Hohn. 1997. Alternative structures of the cauliflower mosaic virus 35 S RNA leader: Implications for viral expression and replication. J. Mol. Biol. 267:1075-1088[CrossRef][Medline]. |
| 196. | Hemmings-Mieszczak, M., G. Steger, and T. Hohn. 1998. Regulation of CaMV 35 S RNA translation is mediated by a stable hairpin in the leader. RNA 4:101-111[Abstract]. |
| 197. |
Hentze, M. W.
1997.
eIF4G: a multipurpose ribosome adapter?
Science
275:500-501 |
| 198. | Hershey, J. W. B. 1991. Translational control in mammalian cells. Annu. Rev. Biochem. 60:717-755[CrossRef][Medline]. |
| 199. | Hershey, J. W. B., M. B. Mathews, and N. Sonenberg (ed.). 1996. Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 200. |
Herson, D.,
A. Schmidt,
S. Seal,
A. Marcus, and L. van Vloten-Doting.
1979.
Competitive mRNA translation in an in vitro system from wheat germ.
J. Biol. Chem.
254:8245-8249 |
| 201. |
Hinnebusch, A. G.
1994.
The eIF-2 kinases: regulators of protein synthesis in starvation and stress.
Semin. Cell Biol.
5:417-426[CrossRef][Medline].
|
| 202. | Hinnebusch, A. G. 1996. Translational control of GCN4: gene-specific regulation by phosphorylation of eIF2, p. 199-244. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 203. | Hohn, T., D. I. Dominguez, N. ScharerHernandez, W. Schmidt-Puchta, M. Hemmings-Mieszczak, and J. Fütterer. 1988. Ribosome shunting in eukaryotes: what the viruses tell me., p. 84-95. In A look beyond transcription. |
| 204. |
Honda, M.,
M. R. Beard,
L. H. Ping, and S. M. Lemon.
1999.
A phylogenetically conserved stem-loop structure at the 5' border of the internal ribosome entry site of hepatitis C virus is required for cap-independent viral translation.
J. Virol.
73:1165-1174 |
| 205. | Honda, M., E. A. Brown, and S. M. Lemon. 1996. Stability of a stem-loop involving the initiator AUG controls the efficiency of internal initiation of translation on hepatitis C virus RNA. RNA 2:955-968[Abstract]. |
| 206. | Honda, M., L. H. Ping, R. A. Rijnbrand, E. Amphlett, B. Clarke, D. Rowlands, and S. M. Lemon. 1996. Structural requirements for initiation of translation by internal ribosome entry within genome-length hepatitis C virus RNA. Virology 222:31-42[CrossRef][Medline]. |
| 207. |
Honda, M.,
R. Rijnbrand,
G. Abell,
D. Kim, and S. M. Lemon.
1999.
Natural variation in translational activities of the 5' nontranslated RNAs of hepatitis C virus genotypes 1a and 1b: evidence for a long-range RNA-RNA interaction outside of the internal ribosome entry site.
J. Virol.
73:4941-4951 |
| 208. | Hoofnagle, J. H. 1994. Therapy of acute and chronic viral hepatitis. Adv. Intern. Med. 39:241-275[Medline]. |
| 209. | Houser-Scott, F., P. Ansel-McKinney, J. M. Cai, and L. Gehrke. 1997. In vitro genetic selection analysis of alfalfa mosaic virus coat protein binding to 3'-terminal AUGC repeats in the viral RNAs. J. Virol. 71:2310-2319[Abstract]. |
| 210. | Huang, J., and R. J. Schneider. 1991. Adenovirus inhibition of cellular protein synthesis involves inactivation of cap-binding protein. Cell 65:271-280[CrossRef][Medline]. |
| 211. | Hwang, L. H., C. L. Hsieh, A. Yen, Y. L. Chung, and D. S. Chen. 1998. Involvement of the 5' proximal coding sequences of hepatitis C virus with internal initiation of viral translation. Biochem. Biophys. Res. Commun. 252:455-460[CrossRef][Medline]. |
| 212. |
Icely, P. L.,
P. Gros,
J. J. M. Bergeron,
A. Devault,
D. E. H. Afar, and J. C. Bell.
1991.
TIK, a novel serine/threonine kinase, is recognized by antibodies directed against phosphotyrosine.
J. Biol. Chem.
266:16073-16077 |
| 213. | Iino, S., K. Hino, and K. Yasuda. 1994. Current state of interferon therapy for chronic hepatitis C. Intervirology 37:87-100[Medline]. |
| 214. | Imataka, H., A. Gradi, and N. Sonenberg. 1998. A newly identified N-terminal amino acid sequence of human eIF4G binds poly(A)-binding protein and functions in poly(A)-dependent translation. EMBO J. 17:7480-7489[CrossRef][Medline]. |
| 215. |
Inglis, S. C.
1982.
Inhibition of host protein synthesis and degradation of cellular mRNAs during infection by influenza and Herpes simplex virus.
Mol. Cell. Biol.
2:1644-1648 |
| 216. | Ito, T., and M. M. C. Lai. 1997. Determination of the secondary structure of and cellular protein binding to the 3' untranslated region of the hepatitis C virus RNA genome. J. Virol. 71:8698-8706[Abstract]. |
| 217. | Ito, T., and M. M. C. Lai. 1999. An internal polyprimidine-tract-binding protein-binding site in the hepatitis C virus RNA attenuates translation, which is relieved by the 3'-untranslated region. Virology 254:288-296[CrossRef][Medline]. |
| 218. |
Ito, T.,
S. M. Tahara, and M. C. Lai.
1998.
The 3'-untranslated region of hepatitis C virus RNA enhances translation from an internal ribosomal entry site.
J. Virol.
72:8789-8796 |
| 219. | Jacks, T., H. D. Madhani, F. R. Masiarz, and H. E. Varmus. 1988. Signals for ribosomal frameshifting in the Rous sarcoma virus gag-pol region. Cell 55:447-458[CrossRef][Medline]. |
| 220. |
Jacks, T., and H. E. Varmus.
1985.
Expression of the Rous sarcoma virus pol gene by ribosomal frameshifting.
Science
230:1237-1242 |
| 221. | Jackson, R. J. 1996. A comparative view of initiation site selection mechanisms, p. 71-112. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 222. | Jackson, R. J., M. T. Howell, and A. Kaminski. 1990. The novel mechanism of initiation of picornavirus RNA translation. Trends Biochem. Sci. 15:477-483[CrossRef][Medline]. |
| 223. | Jackson, R. J., and A. Kaminski. 1995. Internal initiation of translation in eukaryotes: the picornavirus paradigm and beyond. RNA 1:985-1000[Medline]. |
| 224. |
Jacobs, B. L., and R. E. Ferguson.
1991.
The Lang strain of reovirus serotype 1 and the Dearing strain of reovirus serotype 3 differ in their sensitivities to beta interferon.
J. Virol.
65:5102-5104 |
| 225. | Jacobson, A. 1996. Poly(A) metabolism and translation: the closed-loop model, p. 451-480. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 226. |
Jang, S. K.,
M. V. Davies,
R. J. Kaufman, and E. Wimmer.
1989.
Initiation of protein synthesis by internal entry of ribosomes into the 5' nontranslated region of encephalomyocarditis virus RNA in vivo.
J. Virol.
63:1651-1660 |
| 227. |
Jang, S. K.,
H. G. Krausslich,
M. J. H. Nicklin,
A. C. Duke,
A. C. Palmenberg, and E. Wimmer.
1988.
A segment of the 5' nontranslated region of the encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation.
J. Virol.
62:2636-2643 |
| 228. | Jang, S. K., T. V. Pestova, C. U. T. Hellen, G. W. Witherell, and E. Wimmer. 1990. Cap-independent translation of picornavirus RNAs: Structure and function of the internal ribosomal entry site. Enzyme 44:292-309[Medline]. |
| 229. | Jang, S. K., and E. Wimmer. 1990. Cap-independent translation of encephalomyocarditis virus RNA: structural requirements of the internal ribosome entry site and involvement of a 57kD RNA binding protein. Genes Dev. 4:1560-1572[Abstract]. |
| 230. | Jaramillo, M. L., N. Abraham, and J. C. Bell. 1995. The interferon system: a review with emphasis on the role of PKR in growth control. Cancer Investig. 13:327-338[Medline]. |
| 231. |
Jefferies, H. B., and G. Thomas.
1994.
Elongation factor-1 alpha mRNA is selectively translated following mitogenic stimulation.
J. Biol. Chem.
269:4367-4372 |
| 232. | Jefferies, H. B. J., and G. Thomas. 1996. Ribosomal protein S6 phosphorylation and signal transduction, p. 389-409. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 233. |
Joachims, M.,
P. C. Van-Breugel, and R. E. Lloyd.
1999.
Cleavage of poly(A)-binding protein by enterovirus proteases concurrent with inhibition of translation in vitro.
J. Virol.
73:718-727 |
| 234. |
Johannes, G.,
M. S. Carter,
M. B. Eisen,
P. O. Brown, and P. Sarnow.
1999.
Identification of eukaryotic mRNAs that are translated at reduced cap binding complex eIF4F concentrations using cDNA microarray.
Proc. Natl. Acad. Sci. USA
96:13118-13123 |
| 235. | Johannes, G., and P. Sarnow. 1998. Cap-independent polysomal association of natural mRNAs encoding c-myc, BiP, and eIF4G conferred by internal ribosome entry sites. RNA 4:1500-1513[Abstract]. |
| 236. | Johnston, M. 1998. Gene chips: array of hope for understanding gene regulation. Curr. Biol. 8:R171-R174[CrossRef][Medline]. |
| 237. |
Joshi, B.,
A.-L. Cai,
B. D. Keiper,
W. B. Minich,
R. Mendez,
C. M. Beach,
J. Stepinski,
R. Stolarski,
E. Darzynkiewicz, and R. E. Rhoads.
1995.
Phosphorylation of eukaryotic protein synthesis initiation factor 4E at ser-209.
J. Biol. Chem.
270:14597-14603 |
| 238. | Kaminski, A., S. L. Hunt, J. G. Patton, and R. J. Jackson. 1995. Direct evidence that polypyrimidine tract binding protein (PTB) is essential for internal initiation of translation of encephalomyocarditis virus RNA. RNA 1:924-938[Abstract]. |
| 239. | Kaminski, A., and R. J. Jackson. 1998. The polypyrimidine tract binding protein (PTB) requirement for internal initiation of translation of cardiovirus RNAs is conditional rather than absolute. RNA 4:626-638[Abstract]. |
| 240. |
Kato, N.,
M. Hijikata,
Y. Ootsuyama,
M. Nakagawa,
S. Ohkoshi,
T. Sugimura, and K. Shimotohno.
1990.
Molecular cloning of the human hepatitis C virus genome from Japanese patients with non-A, non-B hepatitis.
Proc. Natl. Acad. Sci. USA
87:9524-9528 |
| 241. | Katze, M. G. 1993. Games viruses play: a strategic initiative against the interferon-induced dsRNA activated 68,000 Mr protein kinase. Semin. Virol. 4:259-268[CrossRef]. |
| 242. | Katze, M. G. 1993. The regulation of the interferon-induced dsRNA activated protein kinase in virus-infected cells by viral and cellular gene products. Semin. Virol. 10:501-508. |
| 243. | Katze, M. G. 1995. The war against the interferon-induced, double-stranded RNA-activated protein kinase: can viruses win? J. Interferon Res. 12:241-248. |
| 244. | Katze, M. G. 1996. Translational control in cells infected with influenza virus and reovirus, p. 607-630. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 245. | Katze, M. G., and M. B. Agy. 1990. Regulation of viral and cellular RNA turnover in cells infected by eukaryotic viruses including HIV-1. Enzyme 44:332-346[Medline]. |
| 246. | Katze, M. G., Y. T. Chen, and R. M. Krug. 1984. Nuclear-cytoplasmic transport and VAI RNA-independent translation of influenza viral messenger RNAs in late adenovirus-infected cells. Cell 37:483-490[CrossRef][Medline]. |
| 247. |
Katze, M. G.,
D. DeCorato, and R. M. Krug.
1986.
Cellular mRNA translation is blocked at both initiation and elongation after infection by influenza virus or adenovirus.
J. Virol.
60:1027-1039 |
| 248. | Katze, M. G., D. DeCorato, B. Safer, J. Galabru, and A. G. Hovanessian. 1987. Adenovirus VAI RNA complexes with the 68,000 Mr protein kinase to regulate its autophosphorylation and activity. EMBO J. 6:689-697[Medline]. |
| 249. |
Katze, M. G.,
B. M. Detjen,
B. Safer, and R. M. Krug.
1986.
Translational control by influenza virus: suppression of the kinase that phosphorylates the alpha subunit of initiation factor eIF-2 and selective translation of influenza viral mRNAs.
Mol. Cell. Biol.
6:1741-1750 |
| 250. |
Katze, M. G., and R. M. Krug.
1984.
Metabolism and expression of RNA polymerase II transcripts in influenza virus infected cells.
Mol. Cell. Biol.
4:2198-2206 |
| 251. | Katze, M. G., and R. M. Krug. 1990. Translational control in influenza virus-infected cells. Enzyme 44:265-277[Medline]. |
| 252. | Katze, M. G., J. Lara, and M. Wambach. 1989. Nontranslated cellular mRNAs are associated with cytoskeletal framework in influenza virus or adenovirus infected cells. Virology 169:312-322[CrossRef][Medline]. |
| 253. |
Katze, M. G.,
J. Tomita,
T. Black,
R. M. Krug,
B. Safer, and A. G. Hovanessian.
1988.
Influenza virus regulates protein synthesis during infection by repressing the autophosphorylation and activity of the cellular 68,000-Mr protein kinase.
J. Virol.
62:3710-3717 |
| 254. |
Katze, M. G.,
M. Wambach,
M.-L. Wong,
M. Garfinkel,
E. Meurs,
K. Chong,
B. R. G. Williams,
A. G. Hovanessian, and G. N. Barber.
1991.
Functional expression and RNA binding analysis of the interferon-induced, double-stranded RNA-activated, 68,000-Mr protein kinase in a cell-free system.
Mol. Cell. Biol.
11:5497-5505 |
| 255. |
Kawagishi-Kobayashi, M.,
J. B. Silverman,
T. L. Ung, and T. E. Dever.
1997.
Regulation of the protein kinase PKR by the vaccinia virus pseudosubstrate inhibitor K3L is dependent on residues conserved between the K3L protein and the PKR substrate eIF-2.
Mol. Cell. Biol.
17:4146-4158[Abstract].
|
| 256. | Kawaguchi, Y., R. Bruni, and B. Roizman. 1997. Interaction of herpes simplex virus 1 alpha regulatory protein ICP0 with elongation factor 1 delta: ICP0 affects translational machinery. J. Virol. 71:1019-1024[Abstract]. |
| 257. |
Kawaguchi, Y.,
C. VanSant, and B. Roizman.
1998.
Eukaryotic elongation factor 1 delta is hyperphosphorylated by the protein kinase encoded by the U(L)13 gene of herpes simplex virus 1.
J. Virol.
72:1731-1736 |
| 258. | Keiper, B. D., W. Gan, and R. E. Rhoads. 1999. Protein synthesis initiation factor 4G. Int. J. Biochem. Cell Biol. 31:37-48[CrossRef][Medline]. |
| 259. |
Kerekatte, V.,
B. D. Keiper,
C. Badorff,
A. L. Cai,
K. U. Knowlton, and R. E. Rhoads.
1999.
Cleavage of poly(A)-binding protein by coxsackievirus 2A protease in vitro and in vivo: another mechanism for host protein synthesis shutoff?
J. Virol.
73:709-717 |
| 260. |
Kimura, T.,
K. Nakayama,
J. Penninger,
M. Kitagawa,
H. Harada,
T. Matsuyama,
N. Tanaka,
R. Kamijo,
J. Vilcek,
T. W. Mak, and T. Taniguchi.
1994.
Involvement of the IRF-1 transcription factor in antiviral responses to interferons.
Science
264:1921-1924 |
| 261. | Kirchhoff, S., A. E. Koromilas, F. Schaper, M. Grashof, N. Sonenberg, and H. Hauser. 1995. IRF-1 induced cell growth inhibition and interferon induction requires the activity of the protein kinase PKR. Oncogene 11:439-445[Medline]. |
| 262. | Knee, R., and P. R. Murphy. 1997. Regulation of gene expression by natural antisense RNA transcripts. Neurochem. Int. 31:379-392[CrossRef][Medline]. |
| 263. | Koff, R. S. 1997. Therapy in chronic hepatitis C: say goodbye to the 6-month interferon regimen. Am. J. Gastroenterol. 91:2072-2074. |
| 264. | Kolupaeva, V. G., C. T. Hellen, and I. N. Shatsky. 1996. Structural analysis of the interaction of the pyrimidine tract-binding protein with the internal ribosomal entry site of encephalomyocarditis virus and foot-and-mouth disease virus RNAs. RNA 2:1199-1212[Abstract]. |
| 265. | Kolykhalov, A. A., S. M. Fienstone, and C. M. Rice. 1996. Identification of a highly conserved sequence element at the 3' terminus of hepatitis C virus genome RNA. J. Virol. 70:3363-3371[Abstract]. |
| 266. |
Koromilas, A. E.,
C. Cantin,
A. W. B. Craig,
R. Jagus,
J. Hiscott, and N. Sonenberg.
1995.
The interferon-inducible protein kinase PKR modulates the transcriptional activation of immunoglobulin kappa gene.
J. Biol. Chem.
270:25426-25434 |
| 267. | Korth, M. J., and M. G. Katze. 1997. mRNA metabolism and cancer, p. 265-280. In J. Harford, and D. Morris (ed.), mRNA metabolism and post-transcriptional gene regulation. Wiley-Liss, Inc., New York, N.Y. |
| 268. | Korth, M. J., C. N. Lyons, M. Wambach, and M. G. Katze. 1996. Cloning, expression, and cellular localization of the oncogenic 58-kDa inhibitor of the RNA-activated human and mouse protein kinase. Gene 170:181-188[CrossRef][Medline]. |
| 269. |
Kozak, M.
1986.
Influences of mRNA secondary structure on initiation by eukaryotic ribosomes.
Proc. Natl. Acad. Sci. USA
83:2850-2854 |
| 270. | Kozak, M. 1986. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukarotic ribosomes. Cell 44:283-292[CrossRef][Medline]. |
| 271. |
Kozak, M.
1991.
An analysis of vertebrate mRNA sequences: intimations of translational control.
J. Cell Biol.
115:887-903 |
| 272. | Kozak, M. 1992. Regulation of translation in eukaryotic systems. Annu. Rev. Cell Biol. 8:197-225[CrossRef]. |
| 273. |
Krausslich, H. G.,
M. J. H. Nicklin,
H. Toyoda,
D. Etchison, and E. Wimmer.
1987.
Poliovirus proteinase 2A induces cleavage of eukaryotic initiation factor 4F polypeptide P220.
J. Virol.
61:2711-2718 |
| 274. | Krug, R. M., F. Alonso-Caplen, I. Julkunen, and M. G. Katze. 1989. Expression and replication of the influenza virus genome, p. 89-152. In R. M. Krug (ed.), Influenza viruses. Plenum Publishing Corp., New York, N.Y. |
| 275. |
Krug, R. M.,
M. Shaw,
B. Broni,
G. Shapiro, and O. Haller.
1985.
Inhibition of influenza viral messenger RNA synthesis in cells expressing the interferon-induced Mx gene product.
J. Virol.
56:201-206 |
| 276. | Kruys, V., and G. Huez. 1994. Translational control of cytokine expression by 3' UA-rich sequences. Biochimie 76:862-866[CrossRef][Medline]. |
| 277. |
Kumar, A.,
Y.-L. Yang,
V. Flati,
S. Der,
S. Kadereit,
A. Deb,
J. Haque,
L. Reis,
C. Weissmann, and B. R. G. Williams.
1997.
Deficient cytokine signaling in mouse embryo fibroblasts with a targeted deletion in the PKR gene: role of IRF-1 and NF-B.
EMBO J.
16:406-416[CrossRef][Medline].
|
| 278. |
Kurosaki, M.,
N. Enomoto,
T. Murakami,
I. Sakuma,
Y. Asahina,
C. Yamamoto,
T. Ikeda,
S. Tozuka,
N. Izumi,
F. Marumo, and C. Sato.
1997.
Analysis of genotypes and amino acid residues 2209 to 2248 of the NS5A region of hepatitis C virus in relation to the response to interferon- therapy.
Hepatology
25:750-753[CrossRef][Medline].
|
| 279. |
Kwong, A. D., and N. Frenkel.
1987.
Herpes simplex virus-infected cells contain a function(s) that destabilizes both host and viral mRNAs.
Proc. Natl. Acad. Sci. USA
84:1926-1930 |
| 280. |
Kwong, A. D.,
J. A. Kruper, and N. Frenkel.
1988.
Herpes simplex virus virion host shutoff function.
J. Virol.
62:912-921 |
| 281. | Laine, R. O., R. G. Hutson, and M. S. Kilberg. 1996. Eukaryotic gene expression: metabolite control by amino acids. Prog. Nucleic Acid Res. Mol. Biol. 53:219-248[Medline]. |
| 282. | Lam, Q., C. A. Smibert, K. E. Koop, C. Lavery, J. P. Capone, S. P. Weinheimer, and J. R. Smiley. 1996. Herpes simplex virus VP16 rescues viral mRNA from destruction by the virion host shutoff function. EMBO J. 15:2575-2581[Medline]. |
| 283. |
Lamphear, B. J.,
R. Kirchweger,
T. Skern, and R. E. Rhoads.
1995.
Mapping of functional domains in eukaryotic protein synthesis initiation factor 4G (eIF4G) with picornaviral proteases. Implications for cap-dependent and cap-independent translational initiation.
J. Biol. Chem.
270:21975-21983 |
| 284. |
Langland, J. O., and B. L. Jacobs.
1992.
Cytosolic double-stranded RNA-dependent protein kinase is likely a dimer of partially phosphorylated Mr 66,000 subunits.
J. Biol. Chem.
267:10729-10736 |
| 285. | Laurent, A. M., J. J. Madjar, and A. Greco. 1998. Translational control of viral and host protein synthesis during the course of herpes simplex virus type 1 infection: evidence that initiation of translation is the limiting step. J. Gen. Virol. 79:2765-2775[Abstract]. |
| 286. |
Lawson, T. G.,
B. K. Ray,
J. T. Dodds,
J. A. Grifo,
R. D. Abramson,
W. C. Merrick,
D. F. Betsch,
H. L. Weith, and R. E. Thach.
1986.
Influence of 5' proximal secondary structure on the translational efficiency of eukaryotic mRNAs and on their interaction with initiation factors.
J. Biol. Chem.
261:13979-13989 |
| 287. | Lazaris-Karatzas, A., K. S. Mantine, and N. Sonenberg. 1990. Malignant transformation by a eukaryotic initiation factor subunit that binds to mRNA 5' cap. Nature 345:544-547[CrossRef][Medline]. |
| 288. | Lazarowitz, S. G., R. W. Compans, and P. W. Choppin. 1971. Influenza virus structural and nonstructural proteins in infected cells and their plasma membrane. Virology 46:830-843[CrossRef][Medline]. |
| 289. |
Le, H.,
R. L. Tanguay,
M. L. Balasta,
C. C. Wei,
K. S. Browning,
A. M. Metz,
D. J. Goss, and D. R. Gallie.
1997.
Translation initiation factors eIF-iso4G and eIF-4B interact with the poly(A)-binding protein and increase its RNA binding activity.
J. Biol. Chem.
272:16247-16255 |
| 290. | Le, S. Y., A. Siddiqui, and J. V. Maizel. 1996. A common structural core in the internal ribosome entry sites of picornavirus, hepatitis C virus, and pestivirus. Virus Genes 12:135-147[CrossRef][Medline]. |
| 291. |
Lee, J. H.,
S. K. Choi,
A. Roll-Mecak,
S. K. Burley, and T. E. Dever.
1999.
Universal conservation in translation initiation revealed by human and archaeal homologs of bacterial translation initiation factor IF2.
Proc. Natl. Acad. Sci. USA
96:4342-4347 |
| 292. |
Lee, K. A. W., and N. Sonenberg.
1982.
Inactivation of cap-binding proteins accompanies the shut-off of host protein synthesis by poliovirus.
Proc. Natl. Acad. Sci. USA
79:3447-3451 |
| 293. | Lee, S. B., and M. Esteban. 1994. The interferon-induced double-stranded RNA-activated protein kinase induces apoptosis. Virology 199:491-496[CrossRef][Medline]. |
| 294. |
Lee, T. G.,
N. Tang,
S. Thompson,
J. Miller, and M. G. Katze.
1994.
The 58,000-dalton cellular inhibitor of the interferon-induced double-stranded RNA-activated protein kinase (PKR) is a member of the tetratricopeptide repeat family of proteins.
Mol. Cell. Biol.
14:2331-2342 |
| 295. |
Lee, T. G.,
J. Tomita,
A. G. Hovanessian, and M. G. Katze.
1990.
Purification and partial characterization of a cellular inhibitor of the interferon-induced protein kinase of Mr 68,000 from influenza virus-infected cells.
Proc. Natl. Acad. Sci. USA
87:6208-6212 |
| 296. |
Lee, T. G.,
J. Tomita,
A. G. Hovanessian, and M. G. Katze.
1992.
Characterization and regulation of the 58,000-dalton cellular inhibitor of the interferon-induced, dsRNA-activated protein kinase.
J. Biol. Chem.
267:14238-14243 |
| 297. |
Lenardo, M. J.,
C.-M. Fan,
T. Maniatis, and D. Baltimore.
1989.
The involvement of NF-B in -interferon gene regulation reveals its role as widely inducible mediator of signal transduction.
Cell
57:287-294[CrossRef][Medline].
|
| 298. | Lenk, R., and S. Penman. 1979. The cytoskeleton framework and poliovirus metabolism. Cell 16:289-301[CrossRef][Medline]. |
| 299. | Lennon, G. G., and H. Lehrach. 1991. Hybridization analyses of arrayed cDNA libraries. Trends Genet. 7:314-317[Medline]. |
| 300. | Lewis, J. D., and E. Izaurralde. 1997. The role of the cap structure in RNA processing and nuclear export. Eur. J. Biochem. 247:462-469. |
| 301. |
Li, J., and R. A. Petryshyn.
1991.
Activation of the double-stranded RNA-dependent eIF-2 kinase by cellular RNA from 3T3-F442A cells.
Eur. J. Biochem.
195:41-48[Abstract].
|
| 302. | Li, X., and S. L. Rhode, III. 1993. The parvovirus H-1 NS2 protein affects viral gene expression through sequences in the 3' untranslated region. Virology 194:10-19[CrossRef][Medline]. |
| 303. |
Licis, N.,
J. Vanduin,
Z. Baklava, and V. Berzins.
1998.
Long-range translational coupling in single-stranded RNA bacteriophages: an evolutionary analysis.
Nucleic Acids Res.
26:3242-3246 |
| 304. | Lin, C. H., and J. G. Patton. 1995. Regulation of alternative 3' splice site selection by constitutive splicing factors. RNA 1:234-245[Abstract]. |
| 305. |
Lin, T.-A.,
X. Kong,
T. A. J. Haystead,
A. Pause,
G. Belsham,
N. Sonenberg, and J. C. Lawrence, Jr.
1994.
PHAS-I as a link between mitogen-activated protein kinase and translation initiation.
Science
266:653-656 |
| 306. | Lindquist, S., and R. Petersen. 1990. Selective translation and degradation of heat-shock messenger RNAs in Drosophila. Enzyme 44:147-166[Medline]. |
| 307. |
Lloyd, R. M., and A. J. Shatkin.
1992.
Translational stimulation by reovirus polypeptide 3: substitution for VAI RNA and inhibition of phosphorylation of the subunit of eukaryotic initiation factor 2.
J. Virol.
66:6878-6884 |
| 308. | Lockhart, D. J., H. Dong, M. C. Byrne, M. T. Follettie, M. V. Gallo, M. S. Chee, M. Mittmann, C. Wang, M. Kobayashi, H. Horton, and E. L. Brown. 1996. Expression monitoring by hybridization to high-density oligonucleotide arrays. Nat. Biotechnol. 14:1675-1680[CrossRef][Medline]. |
| 309. |
Lodish, H. F., and M. Porter.
1980.
Translational control of protein synthesis after infection by vesicular stomatitis virus.
J. Virol.
36:719-733 |
| 310. |
Logan, J., and T. Shenk.
1984.
Adenovirus tripartite leader sequence enhances translation of mRNAs late after infection.
Proc. Natl. Acad. Sci. USA
81:3655-3659 |
| 311. | Lu, Y., X.-Y. Qian, and R. M. Krug. 1994. The influenza virus NS1 protein: a novel inhibitor of pre-mRNA splicing. Genes Dev. 8:1817-1828[Abstract]. |
| 312. | Lu, Y., M. Wambach, M. G. Katze, and R. M. Krug. 1995. Binding of the influenza virus NS1 protein to double-stranded RNA inhibits the activation of the protein kinase that phosphorylates the eIF-2 translation initiation factor. Virology 214:222-228[CrossRef][Medline]. |
| 313. | Maciejewski, J. P., and S. C. St. Jeor. 1999. Human cytomegalovirus infection of human hematopoietic progenitor cells. Leuk. Lymphoma 33:1-13[Medline]. |
| 314. | Maitra, R. K., N. McMillan, S. Desai, J. McSwiggen, A. G. Hovanessian, G. Sen, B. R. G. Williams, and R. H. Silverman. 1994. HIV-1 TAR RNA has an intrinsic ability to activate interferon-inducible enzymes. Virology 204:823-827[CrossRef][Medline]. |
| 315. |
Makkinje, A.,
H. Xiong,
M. Li, and Z. Damuni.
1995.
Phosphorylation of eukaryotic protein synthesis initiation factor 4E by insulin-stimulated protamine kinase.
J. Biol. Chem.
270:14824-14826 |
| 316. |
Manche, L.,
S. R. Green,
C. Schmedt, and M. B. Mathews.
1992.
Interactions between dsRNA regulators and the protein kinase, DAI.
Mol. Cell. Biol.
12:5238-5248 |
| 317. |
Maran, A.,
R. K. Maitra,
A. Kumar,
B. Dong,
W. Xiao,
G. Li,
B. R. G. Williams,
P. F. Torrence, and R. H. Silverman.
1994.
Blockage of NF-B signaling by selective ablation of an mRNA target by 2-5A antisense chimeras.
Science
265:789-792 |
| 318. | Marcotrigiano, J., A. C. Gingras, N. Sonenberg, and S. K. Burley. 1997. Cocrystal structure of the messenger RNA 5' cap-binding protein (eIF4E) bound to 7-methyl-GDP. Cell 89:951-961[CrossRef][Medline]. |
| 319. | Marczinke, B., R. Fisher, M. Vidakovic, A. J. Bloys, and I. Brierley. 1998. Secondary structure and mutational analysis of the ribosomal frameshift signal of Rous sarcoma virus. J. Mol. Biol. 284:205-225[CrossRef][Medline]. |
| 320. | Mathews, M. B. 1990. Control of translation in adenovirus-infected cells. Enzyme 44:250-264[Medline]. |
| 321. | Mathews, M. B. 1993. Viral evasion of cellular defense mechanisms: regulation of the protein kinase DAI by RNA effectors. Semin. Virol. 4:247-258[CrossRef]. |
| 322. | Mathews, M. B. 1996. Interactions between viruses and the cellular machinery for protein synthesis, p. 505-548. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 323. |
Mathews, M. B., and T. Shenk.
1991.
Adenovirus virus-associated RNA and translation control.
J. Virol.
65:5657-5662 |
| 324. |
McCarthy, J. E. G.
1998.
Postranscriptional control of gene expression in yeast.
Microbiol. Mol. Biol. Rev.
62:1492-1553 |
| 325. |
McCormack, S. J., and C. E. Samuel.
1995.
Mechanism of interferon action: RNA-binding activity of full-length and R-domain forms of the RNA-dependent protein kinase PKR determination of KD values for VA1 and TAR RNAs.
Virology
206:511-519[CrossRef][Medline].
|
| 326. |
McHutchison, J. G.,
S. C. Gordon,
E. R. Schiff,
M. L. Shiffman,
W. M. Lee,
Z. D. Goodman,
M. H. Ling,
S. Cort, and J. K. Albrecht for the Interferon Interventional Therapy Group.
1998.
Interferon alfa-2b alone or in combination with ribavirin as initial treatment for chronic hepatitis C.
N. Engl. J. Med.
339:1485-1492 |
| 327. | McMillan, N. A. J., R. F. Chun, D. P. Siderovski, J. Galabru, W. M. Toone, C. E. Samuel, T. W. Mak, A. G. Hovanessian, K.-T. Jeang, and B. R. G. Williams. 1995. HIV-1 Tat directly interacts with the interferon-induced, double-stranded RNA-dependent kinase, PKR. Virology 213:413-424[CrossRef][Medline]. |
| 328. | Meerovitch, K., J. Pelletier, and N. Sonenberg. 1989. A cellular protein that binds to the 5'-noncoding region of poliovirus RNA: implications for internal translation initiation. Genes Dev. 13:1026-1034. |
| 329. | Meerovitch, K., and N. Sonenberg. 1993. Internal initiation of picornavirus RNA translation. Semin. Virol. 4:504.1-504.11. |
| 330. |
Meerovitch, K.,
Y. V. Svitkin,
H. S. Lee,
F. Lejbkowicz,
D. J. Kenan,
E. K. L. Chan,
V. I. Agol,
J. D. Keene, and N. Sonenberg.
1993.
La autoantigen enhances and corrects aberrant translation of poliovirus RNA in reticulocyte lysate.
J. Virol.
67:3798-3807 |
| 331. | Mellits, K. H., M. Kostura, and M. B. Mathews. 1990. Interaction of adenovirus VA RNA1 with the protein kinase DAI: nonequivalence of binding and function. Cell 61:843-852[CrossRef][Medline]. |
| 332. |
Melville, M. W.,
W. J. Hansen,
B. C. Freeman,
W. J. Welch, and M. G. Katze.
1997.
The molecular chaperone hsp40 regulates the activity of P58IPK, the cellular inhibitor of PKR.
Proc. Natl. Acad. Sci. USA
94:97-102 |
| 333. |
Melville, M. W.,
S.-L. Tan,
M. Wambach,
J. Song,
R. I. Morimoto, and M. G. Katze.
1999.
The cellular inhibitor of the PKR protein kinase, P58IPK, is an influenza virus-activated co-chaperone that modulates heat shock protein 70 activity.
J. Biol. Chem.
274:3797-3803 |
| 334. | Meraz, M. A., J. M. White, K. C. F. Sheehan, E. A. Bach, S. J. Rodig, A. S. Dighe, D. H. Kaplan, J. K. Riley, A. C. Greenlund, D. Campbell, K. Carver-Moore, R. N. DuBois, R. Clark, M. Aguet, and R. D. Schreiber. 1996. Targeted disruption of the Stat1 gene in mice reveals unexpected physiologic specificity in the JAK-STAT signaling pathway. Cell 84:431-442[CrossRef][Medline]. |
| 335. | Merrick, W. C., and J. W. B. Hershey. 1996. The pathway and mechanism of eukaryotic protein synthesis, p. 31-70. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 336. | Meurs, E., K. L. Chong, J. Galabru, N. Thomas, I. Kerr, B. R. G. Williams, and A. G. Hovanessian. 1990. Molecular cloning and characterization of the human double-stranded RNA-activated protein kinase induced by interferon. Cell 62:379-390[CrossRef][Medline]. |
| 337. |
Meurs, E. F.,
J. Galabru,
G. N. Barber,
M. G. Katze, and A. G. Hovanessian.
1993.
Tumor suppressor function of the interferon-induced double-stranded RNA-activated protein kinase.
Proc. Natl. Acad. Sci. USA
90:232-236 |
| 338. | Mohr, I., and Y. Gluzman. 1996. A herpesvirus genetic element which affects translation in the absence of the viral GADD34 function. EMBO J. 15:4759-4766[Medline]. |
| 339. | Morley, S. J. 1997. Signalling through either the p38 or ERK mitogen-activated protein (MAP) kinase pathway is obligatory for phorbol ester and T cell receptor complex (TCR-CD3)-stimulated phosphorylation of initiation factor (eIF) 4E in Jurkat T cells. FEBS Lett. 418:327-332[CrossRef][Medline]. |
| 340. | Morley, S. J., P. S. Curtis, and V. M. Pain. 1997. eIF4G: Translation's mystery factor begins to yield its secrets. RNA 3:1085-1104[Medline]. |
| 341. |
Morley, S. J., and L. McKendrick.
1997.
Involvement of stress-activated protein kinase and p38/RK mitogen-activated protein kinase signaling pathways in the enhanced phosphorylation of initiation factor 4E in NIH 3T3 cells.
J. Biol. Chem.
272:17887-17893 |
| 342. | Mueller, P. P., and A. G. Hinnebusch. 1986. Multiple upstream AUG codons mediate translational control of GCN4. Cell 45:201-207[CrossRef][Medline]. |
| 343. |
Mulvey, M.,
J. Poppers,
A. Ladd, and I. Mohr.
1999.
A herpesvirus ribosome-associated, RNA-binding protein confers a growth advantage upon mutants deficient in a GADD34-related function.
J. Virol.
73:3375-3385 |
| 344. |
Mundschau, L. J., and D. V. Faller.
1995.
Platelet-derived growth factor signal transduction through the interferon-inducible kinase PKR.
J. Biol. Chem.
270:3100-3106 |
| 345. | Munemitsu, S. M., and C. E. Samuel. 1988. Biosynthesis of reovirus-specified polypeptides: effect of point mutation of the sequences flanking the 5'-proximal AUG initiator codons of the reovirus S1 and S4 genes on the efficiency of mRNA translation. Virology 163:643-646[CrossRef][Medline]. |
| 346. |
Müller, U.,
U. Steinhoff,
L. F. L. Reis,
S. Hemmi,
J. Pavlovic,
R. M. Zinkernagel, and M. Aguet.
1994.
Functional role of type I and type II interferons in antiviral defense.
Science
264:1918-1921 |
| 347. |
Nanbru, C.,
I. Lafon,
S. Audigier,
M. C. Gensac,
S. Vagner,
G. Huez, and A. C. Prats.
1997.
Alternative translation of the proto-oncogene c-myc by an internal ribosome entry site.
J. Biol. Chem.
272:32061-32066 |
| 348. |
Neumann, A. U.,
N. P. Lam,
H. Dahari,
D. R. Gretch,
T. E. Wiley,
T. J. Layden, and A. S. Perelson.
1999.
Hepatitis C virus dynamics in vivo and the antiviral efficacy of interferon- therapy.
Science
282:103-107 |
| 349. | Ng, T. I., Y. E. Chang, and B. Roizman. 1997. Infected cell protein 22 of herpes simplex virus 1 regulates the expression of virion host shutoff gene U(L)41. Virology 234:226-234[CrossRef][Medline]. |
| 350. | Nguyen, C., D. Rocha, S. Granjeaud, M. Baldit, K. Bernard, P. Naquet, and B. R. Jordan. 1995. Differential gene expression in the murine thymus assayed by quantitative hybridization of arrayed cDNA clones. Genomics 20:207-216. |
| 351. |
Nicholson, R.,
J. Pelletier,
S.-Y. Le, and N. Sonenberg.
1991.
Structural and functional analysis of the ribosone landing pad of poliovirus type 2in vivo translation studies.
J. Virol.
65:5886-5894 |
| 352. |
Nishioka, Y., and S. Silverstein.
1977.
Degradation of cellular mRNA during infection by herpes simplex virus.
Proc. Natl. Acad. Sci. USA
74:2370-2374 |
| 353. |
O'Brien, V.
1998.
Viruses and apoptosis.
J. Gen. Virol.
79:1833-1845 |
| 354. | Ohlmann, T., M. Rau, V. M. Pain, and S. J. Morley. 1996. The C-terminal domain of eukaryotic protein synthesis initiation factor (eIF) 4G is sufficient to support cap-independent translation in the absence of eIF4E. EMBO J. 15:1371-1382[Medline]. |
| 355. | Ohsawa, M., N. Shingu, H. Miwa, H. Yoshihara, M. Kubo, H. Tsukuma, H. Teshima, M. Hashimoto, and K. Aozasa. 1999. Risk of non-Hodgkin's lymphoma in patients with hepatitis C virus infection. Int. J. Cancer 80:237-239[CrossRef][Medline]. |
| 356. | Okuda, K. 1998. Hepatitis C and hepatocellular carcinoma. J. Gastroenterol. Hepatol. 13:294-298[Medline]. |
| 357. | Ortega, L. G., M. D. McCotter, G. L. Henry, S. J. McCormack, D. C. Thomis, and C. E. Samuel. 1996. Mechanism of interferon action. Biochemical and genetic evidence for the intermolecular association of the RNA-dependent protein kinase PKR from human cells. Virology 215:31-39[CrossRef][Medline]. |
| 358. | Overton, H., D. McMillan, L. Hope, and P. Wong-Kai-In. 1994. Production of host shutoff-defective mutants of herpes simplex virus type 1 by inactivation of the UL13 gene. Virology 202:97-106[CrossRef][Medline]. |
| 359. | Pain, V. M. 1996. Initiation of protein synthesis in eukaryotic cells. Eur. J. Biochem. 236:747-771[Abstract]. |
| 360. |
Pardigon, N., and J. H. Strauss.
1992.
Cellular proteins bind to the 3' end of Sindbis virus minus-strand RNA.
J. Virol.
66:1007-1015 |
| 361. |
Park, Y.-W., and M. G. Katze.
1995.
Translational control by influenza virus: identification of cis-acting sequences and trans-acting factors which may regulate selective viral mRNA translation.
J. Biol. Chem.
270:28433-28439 |
| 362. |
Park, Y.-W.,
J. Wilusz, and M. G. Katze.
1999.
Regulation of eukaryotic protein synthesis: selective influenza viral mRNA translation is mediated by the cellular RNA-binding protein GRSF-1.
Proc. Natl. Acad. Sci. USA
96:6694-6699 |
| 363. |
Patel, R., and G. C. Sen.
1992.
Identification of the double stranded RNA-binding domain of the human interferon-inducible protein kinase.
J. Biol. Chem.
267:7871-7876 |
| 364. |
Patel, R. C.,
P. Stanton,
N. M. J. McMillan,
B. R. G. Williams, and G. C. Sen.
1995.
The interferon-inducible double-stranded RNA-activated protein kinase self-associates in vitro and in vivo.
Proc. Natl. Acad. Sci. USA
92:8283-8287 |
| 365. |
Patel, R. C.,
P. Stanton, and G. C. Sen.
1996.
Specific mutations near the amino terminus of double-stranded RNA-dependent protein kinase (PKR) differentially affect its double-stranded RNA binding and dimerization properties.
J. Biol. Chem.
271:25657-25663 |
| 366. | Patel, R. C., and G. C. Sen. 1998. PACT, a protein activator of the interferon-induced protein kinase, PKR. EMBO J. 17:4379-4390[CrossRef][Medline]. |
| 367. | Pause, A., G. J. Belsham, A. C. Gingras, O. Donzé, T. A. Lin, J. C. Lawrence, and N. Sonenberg. 1994. Insulin-dependent stimulation of protein synthesis by phosphorylation of a regulator of 5'-cap function. Nature 371:762-767[CrossRef][Medline]. |
| 368. | Pavlakis, G. N., and B. K. Felber. 1990. Regulation of expression of human immunodeficiency virus. New Biol. 2:20-31[Medline]. |
| 369. | Pavlovic, J., and P. Staehli. 1991. The antiviral potentials of Mx proteins. J. Interferon Res. 11:215-219[Medline]. |
| 370. | Pelletier, J., and N. Sonenberg. 1988. Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature 334:320-325[CrossRef][Medline]. |
| 371. |
Pelletier, J., and N. Sonenberg.
1989.
Internal binding of eukaryotic ribosomes on poliovirus RNA: translation in HeLa cell extracts.
J. Virol.
63:441-444 |
| 372. | Pestova, T. V., C. T. Hellen, and I. N. Shatsky. 1996. Canonical eukaryotic initiation factors determine initiation of translation by internal ribosomal entry. Mol. Cell Biol. 16:6859-6869[Abstract]. |
| 373. |
Petryshyn, R. A.,
A. G. Ferrenz, and J. Li.
1997.
Characterization and mapping of the double-stranded regions involved in activation of PKR within a cellular RNA from 3T3-F442A cells.
Nucleic Acids Res.
25:2672-2678 |
| 374. | Petska, S., J. A. Langer, K. Zoon, and C. E. Samuel. 1987. Interferons and their actions. Annu. Rev. Biochem. 56:727-777[CrossRef][Medline]. |
| 375. | Pietu, G., O. Alibert, V. Guichard, B. Lamy, F. Bois, E. Leroy, R. Mariage-Samson, R. Houlgatee, P. Soularue, and C. Auffray. 1996. Novel gene transcripts preferentially expressed in human muscles revealed by quantitative hybridization of a high density cDNA array. Genome Res. 6:492-503[Abstract]. |
| 376. | Piron, M., P. Vende, J. Cohen, and D. Poncet. 1998. Rotavirus RNA-binding protein NSP3 interacts with eIF4GI and evicts the poly(A) binding protein from eIF4F. EMBO J. 17:5811-5821[CrossRef][Medline]. |
| 377. | Polyak, S. J., D. M. Paschal, S. Mcardle, M. J. Gale, Jr., D. Moradpour, and D. R. Gretch. 1999. Characterization of the effects of hepatitis C virus nonstructural 5A protein expression in human cell lines and on interferon-sensitive virus replication. Hepatology 29:1262-1271[CrossRef][Medline]. |
| 378. |
Polyak, S. J.,
N. Tang,
M. Wambach,
G. N. Barber, and M. G. Katze.
1996.
The p58 cellular inhibitor complexes with the interferon-induced, double-stranded RNA-dependent protein kinase, PKR, to regulate its autophosphorylation and activity.
J. Biol. Chem.
271:1702-1707 |
| 379. | Poon, A. W., and B. Roizman. 1997. Differentiation of the shutoff of protein synthesis by virion host shutoff and mutant gamma(1)34.5 genes of herpes simplex virus 1. Virology 229:98-105[CrossRef][Medline]. |
| 380. |
Poulin, F.,
A.-C. Gingras,
H. Olsen, and N. Sonenberg.
1998.
4E-BP3, a new member of the eukaryotic initiation factor 4E-binding protein family.
J. Biol. Chem.
273:14002-14007 |
| 381. |
Pratt, G.,
A. R. Galpine,
N. A. Sharp,
S. Palmer, and M. J. Clemens.
1988.
Regulation of in vitro translation by double-stranded RNA in mammalian cell mRNA preparations.
Nucleic Acids Res.
16:3497-3510 |
| 382. | Proud, C. G. 1995. PKR: a new name and new roles. Trends Biochem. Sci. 20:241-246[CrossRef][Medline]. |
| 383. | Pyronnet, S., H. Imataka, A.-C. Gingras, R. Fukunaga, T. Hunter, and N. Sonenberg. 1999. Human eukaryotic translation initiation factor 4G (eIF4G) recruits Mnk1 to phosphorylate eIF4E. EMBO J. 18:270-279[CrossRef][Medline]. |
| 384. |
Qian, Z., and J. Wilusz.
1994.
GRSF-1: a poly(A)+ mRNA binding protein which interacts with a conserved G-rich element.
Nucleic Acids Res.
22:2334-2343 |
| 385. |
Qiu, Y., and R. M. Krug.
1994.
The influenza virus NS1 protein is a poly(A)-binding protein that inhibits nuclear export of mRNAs containing poly(A).
J. Virol.
68:2425-2432 |
| 386. | Rajan, P., S. Swaminathan, J. Zhu, C. N. Cole, G. Barber, M. J. Tevethia, and B. Thimmapaya. 1995. A novel translational regulation function for the simian virus 40 large T-antigen gene. J. Virol. 69:785-795[Abstract]. |
| 387. | Raught, B., and A.-C. Gingras. 1999. eIF4E activity is regulated at multiple levels. Int. J. Biochem. Cell Biol. 31:43-57[CrossRef][Medline]. |
| 388. |
Ray, B. K.,
T. G. Brendler,
S. Adya,
S. Daniels-McQueen,
J. K. Miller,
J. W. B. Hershey,
J. A. Grifo,
W. C. Merrick, and R. E. Thach.
1983.
Role of mRNA competition in regulating translation: further characterization of mRNA discriminatory initiation factors.
Proc. Natl. Acad. Sci. USA
80:663-667 |
| 389. |
Read, G. S.,
B. M. Karr, and K. Knight.
1993.
Isolation of a herpes simplex virus type 1 mutant with a deletion in the virion host shutoff gene and identification of multiple forms of the vhs (UL41) polypeptide.
J. Virol.
67:7149-7160 |
| 390. | Reed, K. E., and C. M. Rice. 2000. Overview of hepatitis C virus genome structure, polyprotein processing, and protein properties. Curr. Top. Microbiol. Immunol. 242:55-84[Medline]. |
| 391. | Reynolds, J. E., A. Kaminski, A. R. Carroll, B. E. Clarke, D. J. Rowlands, and R. J. Jackson. 1996. Internal initiation of translation of hepatitis C virus RNA: the ribosome entry site is at the authentic initiation codon. RNA 2:867-878[Abstract]. |
| 392. | Rhoads, R. E., S. Joshi-Barve, and C. Rinker-Schaeffer. 1993. Mechanism of action and regulation of protein synthesis initiation factor 4E: effects on mRNA discrimination, cellular growth rate, and oncogenesis. Prog. Nucleic Acid Res. 46:183-219[Medline]. |
| 393. | Richter, J. D. 1996. Dynamics of poly(A) addition and removal during development, p. 481-504. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translation control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 394. | Roberts, L. O., R. A. Seamons, and G. J. Belsham. 1998. Recognition of picornavirus internal ribosome entry sites within cells; influence of cellular and viral proteins. RNA 4:520-529[Abstract]. |
| 395. |
Romano, P. R.,
S. R. Green,
G. N. Barber,
M. B. Mathews, and A. G. Hinnebusch.
1995.
Structural requirements for double-stranded RNA binding, dimerization, and activation of the human eIF-2 kinase DAI in Saccharomyces cerevisiae.
Mol. Cell. Biol.
15:365-378[Abstract].
|
| 396. | Rouault, T. A., R. D. Klausner, and J. B. Harford. 1996. Translational control of ferritin, p. 335-362. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 397. |
Roy, S.,
M. G. Katze,
N. T. Parkin,
I. Edery,
A. G. Hovanessian, and N. Sonenberg.
1990.
Control of the interferon-induced 68-kilodalton protein kinase by the HIV-1 tat gene product.
Science
247:1216-1219 |
| 398. | Rueckert, R. R. 1996. Picornaviridae: the viruses and their replication, p. 609-654. In B. N. Fields, D. M. Knipe, R. M. Chanock, M. S. Hirsch, J. L. Melnick, T. P. Monath, and B. Roizman (ed.), Virology, 3rd ed. Raven Press, New York, N.Y. |
| 399. | Ryabova, L. A., A. F. Torgashov, O. V. Kurnasov, M. G. Bubunenko, and A. S. Spirin. 1993. The 3'-terminal intranslated region of alfalfa mosaic virus RNA 4 facilitates the RNA entry into translation in a cell-free system. FEBS Lett. 326:264-266[CrossRef][Medline]. |
| 400. | Sabin, A., and L. Boulger. 1973. History of Sabin attenuated poliovirus oral live vaccine strains. J. Biol. Stand. 1:115-118[CrossRef]. |
| 401. | Sachs, A. B., P. Sarnow, and M. W. Hentze. 1997. Starting at the beginning, middle, and end: translation initiation in eukaryotes. Cell 89:831-838[CrossRef][Medline]. |
| 402. |
Sakamoto, N.,
C.-H. Wu, and G.-Y. Wu.
1996.
Intracellular cleavage of hepatitis C virus RNA and inhibition of viral protein translation by hammerhead ribozymes.
J. Clin. Investig.
98:2720-2728 |
| 403. | Samuel, C. E. 1991. Antiviral actions of interferon interferon-regulated cellular proteins and their surprisingly selective antiviral activities. Virology 183:1-11[CrossRef][Medline]. |
| 404. |
Samuel, C. E.
1993.
The eIF-2 protein kinases, regulators of translation in eukaryotes from yeasts to humans.
J. Biol. Chem.
268:7603-7606 |
| 405. |
Sandri-Goldin, R. M.
1998.
ICP27 mediates HSV RNA export by shuttling through a leucine-rich nuclear export signal and binding viral intronless RNAs through an RGG motif.
Genes Dev.
12:868-879 |
| 406. | Sandri-Goldin, R. M., M. K. Hibbard, and M. A. Hardwicke. 1995. The C-terminal repressor region of herpes simplex virus type 1 ICP27 is required for the redistribution of small nuclear ribonucleoprotein particles and splicing factor SC35; however, these alterations are not sufficient to inhibit host cell splicing. J. Virol. 69:6063-6076[Abstract]. |
| 407. | Schena, M., R. A. Heller, T. P. Theriault, K. Konrad, E. Lachenmeier, and R. W. Davis. 1998. Microarrays: biotechnology's discovery platform for functional genomics. Trends Biotechnol. 16:301-306[CrossRef][Medline]. |
| 408. |
Schena, M.,
D. Shalon,
R. W. Davis, and P. O. Brown.
1995.
Quantitative monitoring of gene expression patterns with a complementary DNA microarray.
Science
270:467-470 |
| 409. | Schiff, L. A. 1998. Reovirus capsid proteins sigma 3 and µ1: interactions that influence viral entry, assembly, and translational control. Curr. Top. Microbiol. Immunol. 233:167-183[Medline]. |
| 410. |
Schmidt-Puchta, W.,
D. I. Dominguez,
D. Lewetag, and T. Hohn.
1997.
Plant ribosome shunting in vitro.
Nucleic Acids Res.
25:2854-2860 |
| 411. | Schneider, R. J. 1996. Adenovirus and vaccinia virus translational control, p. 575-606. In J. W. B. Hershey, M. B. Mathews, and N. Sonenberg (ed.), Translational control. Cold Spring Harbor Laboratory Press, Plainview, N.Y. |
| 412. |
Schwartz, S.,
B. K. Felber,
E.-M. Fenyo, and G. N. Pavlakis.
1990.
Env and Vpu proteins of human immunodeficiency virus type 1 are produced from multiple bicistronic mRNAs.
J. Virol.
64:5448-5456 |
| 413. |
Schwartz, S.,
B. K. Felber, and G. N. Pavlakis.
1992.
Mechanism of translation of monocistronic and human immunodeficiency virus type 1 mRNAs.
Mol. Cell. Biol.
12:207-219 |
| 414. |
Sen, G. C., and P. Lengyel.
1992.
The interferon system: a bird's eye view of its biochemistry.
J. Biol. Chem.
267:5017-5020 |
| 415. | Sen, G. C., and R. M. Ransohoff. 1993. Interferon-induced antiviral actions and their regulation. Adv. Virus Res. 42:57-102[Medline]. |
| 416. | Seydoux, G. 1996. Mechanisms of translational control in early development. Curr. Opin. Genet. Dev. 6:555-561[CrossRef][Medline]. |
| 417. | Sharpe, A. H., and B. N. Fields. 1982. Reovirus inhibition of cellular RNA and protein synthesis: role of the S4 gene. Virology 122:381-391[CrossRef][Medline]. |
| 418. | Sheng, L. X., and I. Tinoco, Jr. 1995. The structure of an RNA pseudoknot that causes efficient frameshifting in mouse mammary tumor virus. J. Mol. Biol. 247:963-978[CrossRef][Medline]. |
| 419. |
Shi, Y.,
K. M. Vattem,
R. Sood,
J. An,
L. Stramm, and R. C. Wek.
1998.
Identification and characterization of pancreatic eukaryotic initiation factor 2 alpha-subunit kinase, PEK, involved in translational control.
Mol. Cell. Biol.
18:7499-7509 |
| 420. | Shors, S. T., E. Beattie, K. Paoletti, J. Tartaglia, and B. L. Jacobs. 1998. Role of the vaccinia virus E3L and K3L gene products in rescue of VSV and EMCV from the effects of IFN-alpha. J. Interferon Cytokine Res. 18:721-729[Medline]. |
| 421. | Silverman, R. H. 1994. Fascination with 2-5A-dependent RNase: a unique enzyme that functions in interferon action. J. Interferon Res. 14:101-104[Medline]. |
| 422. | Silverman, R. H., P. J. Cayley, M. Knight, C. S. Gilbert, and I. M. Kerr. 1982. Control of the ppp(A2'p)nA system in HeLa cells: effects of interferon and virus infection. Eur. J. Biochem. 124:131-138[Abstract]. |
| 423. | Silverman, R. H., and B. R. G. Williams. 1999. Translational control perks up. Nature 397:208-211[CrossRef] |