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PMID: 11333890 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Regulation of mRNA translation and cellular signaling by hepatitis C virus nonstructural protein NS5A.

Journal of virology ·Vol. 75 ·No. 11 ·2001-06-00 ·Pages 5090-8

He Y, Tan SL, Tareen SU, Vijaysri S, Langland JO, Jacobs BL, Katze MG

Abstract

The NS5A nonstructural protein of hepatitis C virus (HCV) has been shown to inhibit the cellular interferon (IFN)-induced protein kinase R (PKR). PKR mediates the host IFN-induced antiviral response at least in part by inhibiting mRNA translation initiation through phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2alpha). We thus examined the effect of NS5A inhibition of PKR on mRNA translation within the context of virus infection by using a recombinant vaccinia virus (VV)-based assay. The VV E3L protein is a potent inhibitor of PKR. Accordingly, infection of IFN-pretreated HeLa S3 cells with an E3L-deficient VV (VVDeltaE3L) resulted in increased phosphorylation levels of both PKR and eIF2alpha. IFN-pretreated cells infected with VV in which the E3L locus was replaced with the NS5A gene (VVNS5A) displayed diminished phosphorylation of PKR and eIF2alpha in a transient manner. We also observed an increase in activation of p38 mitogen-activated protein kinase in IFN-pretreated cells infected with VVDeltaE3L, consistent with reports that p38 lies downstream of the PKR pathway. Furthermore, these cells exhibited increased phosphorylation of the cap-binding initiation factor 4E (eIF4E), which is downstream of the p38 pathway. Importantly, these effects were reduced in cells infected with VVNS5A. NS5A was also found to inhibit activation of the p38-eIF4E pathway in epidermal growth factor-treated cells stably expressing NS5A. NS5A-induced inhibition of eIF2alpha and eIF4E phosphorylation may exert counteracting effects on mRNA translation. Indeed, IFN-pretreated cells infected with VVNS5A exhibited a partial and transient restoration of cellular and viral mRNA translation compared with IFN-pretreated cells infected with VVDeltaE3L. Taken together, these results support the role of NS5A as a PKR inhibitor and suggest a potential mechanism by which HCV might maintain global mRNA translation rate during early virus infection while favoring cap-independent translation of HCV mRNA during late infection.

MeSH Terms
Autoradiography Eukaryotic Initiation Factor-2/metabolism Eukaryotic Initiation Factor-4E Genetic Vectors HeLa Cells Hepacivirus/chemistry,pathogenicity Humans Immunoblotting Interferons/pharmacology Mitogen-Activated Protein Kinases/metabolism Peptide Initiation Factors/metabolism Phosphorylation/drug effects Protein Biosynthesis/drug effects RNA, Messenger/genetics RNA-Binding Proteins/genetics RNA-Dependent RNA Polymerase/physiology Recombinant Proteins/biosynthesis,pharmacology Signal Transduction/drug effects Transfection Vaccinia virus/genetics Viral Nonstructural Proteins/biosynthesis,genetics,pharmacology Viral Proteins/genetics eIF-2 Kinase/antagonists & inhibitors,genetics,metabolism p38 Mitogen-Activated Protein Kinases
Chemicals
E3L protein, Vaccinia virus Eukaryotic Initiation Factor-2 Eukaryotic Initiation Factor-4E NS-5 protein, hepatitis C virus Peptide Initiation Factors RNA, Messenger RNA-Binding Proteins Recombinant Proteins Viral Nonstructural Proteins Viral Proteins Interferons eIF-2 Kinase Mitogen-Activated Protein Kinases p38 Mitogen-Activated Protein Kinases RNA-Dependent RNA Polymerase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
He Y
Department of Microbiology, School of Medicine, University of Washington, Seattle, Washington 98195, USA.
Tan S L
Tareen S U
Vijaysri S
Langland J O
Jacobs B L
Katze M G
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.75.11.5090-5098.2001
Published
2001-06-00
Pages
5090-8
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC114914
Subset
IM
Grants
NCRR NIH HHS · P51 RR000166 · United States
NCRR NIH HHS · RR-00166 · United States
NIAID NIH HHS · AI-22646 · United States
NIAID NIH HHS · R01 AI022646 · United States
NIAID NIH HHS · AI-41629 · United States
External Links
PubMed source DOI: 10.1128/JVI.75.11.5090-5098.2001 PMC Full Text View journal details
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