The Murine Double-Stranded RNA-Dependent Protein Kinase PKR Is Required for Resistance to Vesicular Stomatitis Virus

Stojdl, David F.; Abraham, Ninan; Knowles, Shane; Marius, Ricardo; Brasey, Ann; Lichty, Brian D.; Brown, Earl G.; Sonenberg, Nahum; Bell, John C.

doi:10.1128/jvi.74.20.9580-9585.2000

Cited by 183 publications

(141 citation statements)

References 39 publications

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“…1D, top left). VSV infection did cause an increase in eIF2␣ phosphorylation, as reported previously (11,12). However, this occurred long after the inhibition of host protein synthesis seen in Fig.…”

Section: Eif2␣ Phosphorylation Occurs After Host Protein Synthesis Issupporting

confidence: 46%

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Inhibition of Host and Viral Translation during Vesicular Stomatitis Virus Infection

Connor

Lyles

2005

Journal of Biological Chemistry

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In cells that allow replication of vesicular stomatitis virus (VSV), there are two phases of translation inhibition: an early block of host translation and a later inhibition of viral translation. We investigated the phosphorylation of the ␣ subunit of the eIF2 complex during these two phases of viral infection. In VSV-infected cells, the accumulation of phosphorylated (inactivated) eIF2␣ did not begin until well after host protein synthesis was inhibited, suggesting that it only plays a role in blocking viral translation later after infection. Consistent with this, cells expressing an unphosphorylatable eIF2␣ showed prolonged viral protein synthesis without an effect on host protein synthesis inhibition. Induction of eIF2␣ phosphorylation at early times of viral infection by treatment with thapsigargin showed that virus and host translation are similarly inhibited, demonstrating that viral and host messages are similarly sensitive to eIF2␣ phosphorylation. A recombinant virus that expresses a mutant matrix protein and is defective in the inhibition of host and virus protein synthesis showed an altered phosphorylation of eIF2␣, demonstrating an involvement of viral protein function in inducing this antiviral response. This analysis of eIF2␣ phosphorylation, coupled with earlier findings that the eIF4F complex is modified earlier during VSV infection, supports a temporal/kinetic model of translation control, where at times soon after infection, changes in the eIF4F complex result in the inhibition of host protein synthesis; at later times, inactivation of the eIF2 complex blocks VSV protein synthesis.Many viruses can selectively inhibit host translation and still use the host translation machinery to synthesize their own proteins (for reviews, see Refs. 1-3). This provides two advantages for viral replication. It facilitates the rapid production of viral proteins, and it can also inhibit the production of host antiviral proteins (4). Both factors are important in the race of the virus against the cellular antiviral response. On the other hand, a critical part of the cellular antiviral response is the global inhibition of translation that blocks viral protein synthesis, effectively dampening virus production (5). The inhibition of both host and viral translation are apparent during infection with the prototype rhabdovirus vesicular stomatitis virus (VSV).1 During VSV infection, there is a rapid inhibition of host mRNA translation early after infection. This block of host translation is followed by a later inhibition of viral mRNA translation (6). The experiments presented here address the question of whether these two events reflect differing sensitivities of viral and host mRNAs to a single translation inhibition response or whether there are separate mechanisms controlling each inhibitory event.A substantial body of evidence implicates the phosphorylation of the initiation factor eIF2␣ as a major player in inhibiting viral translation (5). eIF2␣ is a subunit of eIF2, the multiprotein complex that is responsible ...

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Section: Eif2␣ Phosphorylation Occurs After Host Protein Synthesis Issupporting

confidence: 46%

“…More recent studies using cells derived from PKR knock-out mice suggested that the inactivation of eIF2 was caused by eIF2␣ phosphorylation and was primarily a function of PKR (11,12), although a recent publication has demonstrated that there is an additional contribution from pancreatic endoplasmic reticulum kinase (13).…”

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confidence: 99%

Inhibition of Host and Viral Translation during Vesicular Stomatitis Virus Infection

Connor

Lyles

2005

Journal of Biological Chemistry

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“…Each of these pathways affects different stages of the viral life cycle (2,9,13,19,36). However, the virus specificity of each of these pathways is not clearly established and seems dependent on the cell type or the animal model used (32,(37)(38)(39). Thus, additional studies with several other RNA and DNA viruses are needed to precisely clarify the viral specificities of ISG20.…”

Section: Discussionmentioning

confidence: 99%

ISG20, a New Interferon-induced RNase Specific for Single-stranded RNA, Defines an Alternative Antiviral Pathway against RNA Genomic Viruses

Espert

Degols

Gongora

et al. 2003

Journal of Biological Chemistry

197

196

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Interferons (IFNs) encode a family of secreted proteins that provide the front-line defense against viral infections. Their diverse biological actions are thought to be mediated by the products of specific but usually overlapping sets of cellular genes induced in the target cells. We have recently isolated a new human IFN-induced gene that we have termed ISG20, which codes for a 3 to 5 exonuclease with specificity for single-stranded RNA and, to a lesser extent, for DNA. In this report, we demonstrate that ISG20 is involved in the antiviral functions of IFN. In the absence of IFN treatment, ISG20-overexpressing HeLa cells showed resistance to infections by vesicular stomatitis virus (VSV), influenza virus, and encephalomyocarditis virus (three RNA genomic viruses) but not to the DNA genomic adenovirus. ISG20 specifically interfered with VSV mRNA synthesis and protein production while leaving the expression of cellular control genes unaffected. No antiviral effect was observed in cells overexpressing a mutated ISG20 protein defective in exonuclease activity, demonstrating that the antiviral effects were due to the exonuclease activity of ISG20. In addition, the inactive mutant ISG20 protein, which is able to inhibit ISG20 exonuclease activity in vitro, significantly reduced the ability of IFN to block VSV development. Taken together, these data suggested that the antiviral activity of IFN against VSV is partly mediated by ISG20. We thus show that, besides RNase L, ISG20 has an antiviral activity, supporting the idea that it might represent a novel antiviral pathway in the mechanism of IFN action. Interferons (IFNs)1 are a family of multifunctional secreted proteins characterized by their abilities to interfere with virus infection and replication (1, 2). IFNs can indirectly inhibit viral production by reducing the growth of target cells and by stimulating their susceptibility to apoptotic processes (3, 4) or by promoting the recognition and the cytotoxic killing of infected cells by the immune system (5, 6). IFNs also act directly at various steps of the viral multiplication cycle through the products of specific but usually overlapping sets of cellular genes induced in the target cells and involved in RNA and protein metabolism and signaling as well (7,8). Until now, three IFNregulated pathways have been considered to be involved in these processes: the double-stranded RNA-dependent protein kinase R (PKR) (9 -11), the 2-5A/RNase L system (12, 13), and the Mx proteins (14 -16). PKR is a serine/threonine kinase that, after binding to dsRNA, phosphorylates the protein synthesis initiation factor eIF2 and the inhibitor of nuclear factor B (IB), resulting in the inhibition of protein synthesis and specific transcription regulation (reviewed in . RNase L is a dormant cytosolic endoribonuclease that is activated by short oligoadenylates produced by the 2Ј-5Ј oligoadenylate synthetase after viral infection or IFN exposure (reviewed in Refs. 2 and 13). Degradation of viral RNAs and cleavage of cellular 18 S and 28 S rRNA...

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“…In response to viral infection, host dsRNAactivated kinase (PKR) phosphorylates eIF2 in an attempt to block general translation (both cellular and viral) (24)(25)(26)(27). However, most viruses avoid this by expressing products that prevent the activation of PKR in infected cells (reviewed in ref.…”

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confidence: 99%

Inhibition of host translation by virus infection in vivo

Toribio

Ventoso

2010

Proc. Natl. Acad. Sci. U.S.A.

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Infection of cultured cells with lytic animal viruses often results in the selective inhibition of host protein synthesis, whereas viral mRNA is efficiently translated under these circumstances. This phenomenon, known as "shut off," has been well described at the molecular level for some viruses, but there is not yet any direct or indirect evidence supporting the idea that it also should operate in animals infected with viruses. To address this issue, we constructed recombinant Sindbis virus (SV)-expressing reporter mRNA, the translation of which is sensitive or resistant to virus-induced shut off. As found in cultured cells, replication of SV in mouse brain was associated with a strong phosphorylation of eukaryotic initiation factor (eIF2) that prevented translation of reporter mRNA (luciferase and EGFP). Translation of these reporters was restored in vitro, in vivo, and ex vivo when a viral RNA structure, termed downstream hairpin loop, present in viral 26S mRNA, was placed at the 5′ end of reporter mRNAs. By comparing the expression of shut off-sensitive and -resistant reporters, we unequivocally concluded that replication of SV in animal tissues is associated with a profound inhibition of nonviral mRNA translation. A strategy as simple as that followed here might be applicable to other viruses to evaluate their interference on host translation in infected animals.eukaryotic initiation factor-2 phosphorylation | dsRNA-dependent protein kinase | Sindbis

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The Murine Double-Stranded RNA-Dependent Protein Kinase PKR Is Required for Resistance to Vesicular Stomatitis Virus

Cited by 183 publications

References 39 publications

Inhibition of Host and Viral Translation during Vesicular Stomatitis Virus Infection

Inhibition of Host and Viral Translation during Vesicular Stomatitis Virus Infection

ISG20, a New Interferon-induced RNase Specific for Single-stranded RNA, Defines an Alternative Antiviral Pathway against RNA Genomic Viruses

Inhibition of host translation by virus infection in vivo

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