The gamma(1)34.5 protein of herpes simplex virus 1 complexes with protein phosphatase 1alpha 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.
In human cells infected with herpes simplex virus 1 the double-stranded RNA-dependent protein kinase (PKR) is activated but phosphorylation of the alpha subunit of eukaryotic translation initiation factor 2 (eIF-2) and total shutoff of protein synthesis is observed only in cells infected with gamma(1)z34.5- mutants. The carboxyl-terminal 64 aa of gamma(1)34.5 protein are homologous to the corresponding domain of MyD116, the murine growth arrest and DNA damage gene 34 (GADD34) protein and the two domains are functionally interchangeable in infected cells. This report shows that (i) the carboxyl terminus of MyD116 interacts with protein phosphatase 1alpha in yeast, and both MyD116 and gamma(1)34.5 interact with protein phosphatase 1alpha in vitro; (ii) protein synthesis in infected cells is strongly inhibited by okadaic acid, a phosphatase 1 inhibitor; and (iii) the alpha subunit in purified eIF-2 phosphorylated in vitro is specifically dephosphorylated by S10 fractions of wild-type infected cells at a rate 3000 times that of mock-infected cells, whereas the eIF-2alpha-P phosphatase activity of gamma(1)34.5- virus infected cells is lower than that of mock-infected cells. The eIF-2alpha-P phosphatase activities are sensitive to inhibitor 2. In contrast to eIF-2alpha-P phosphatase activity, extracts of mock-infected cells exhibit a 2-fold higher phosphatase activity on [32P]phosphorylase than extracts of infected cells. These results indicate that in infected cells, gamma(1)34.5 interacts with and redirects phosphatase to dephosphorylate eIF-2alpha to enable continued protein synthesis despite the presence of activated PKR. The GADD34 protein may have a similar function in eukaryotic cells. The proposed mechanism for maintenance of protein synthesis in the face of double-stranded RNA accumulation is different from that described for viruses examined to date.
He
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Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, IL 60637, USA.
Gross
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M
Roizman
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D000818:Animals; D000943:Antigens, Differentiation; D018797:Cell Cycle Proteins; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism*; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D006801:Humans; D051379:Mice; D009363:Neoplasm Proteins; D019319:Okadaic Acid / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010765:Phosphorylase Phosphatase / Q000378:metabolism; D010766:Phosphorylation; D014176:Protein Biosynthesis; D054645:Protein Phosphatase 1; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism*; D015003:Yeasts; D019892:eIF-2 Kinase
D000943:Antigens, Differentiation; D018797:Cell Cycle Proteins; D015852:Eukaryotic Initiation Factor-2; D009363:Neoplasm Proteins; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; D010765:Phosphorylase Phosphatase; D054645:Protein Phosphatase 1; D017346:Protein Serine-Threonine Kinases; D011506:Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D019892:eIF-2 Kinase; C066469:gamma 34.5 protein, Human herpesvirus 1; C087519:PPP1R15A protein, human; C513256:Ppp1r15a protein, mouse