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.
Interferon resistance of vaccinia virus is mediated by specific inhibition of phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF-2 alpha) by the double-stranded-RNA-activated (DAI) protein kinase. Vaccinia virus encodes a homolog of eIF-2 alpha, K3L, the deletion of which renders the virus sensitive to interferon treatment. We have studied the mechanism by which this protein product elicits interferon resistance in a transient DNA transfection system designed to evaluate regulators of eIF-2 alpha phosphorylation. In this system, translation of a reporter gene mRNA is inefficient because of eIF-2 phosphorylation mediated by the DAI protein kinase. Cotransfection of the K3L gene enhances translation of the reporter mRNA in this system. The K3L protein inhibits eIF-2 alpha phosphorylation and DAI kinase activation, apparently without being phosphorylated itself. Inhibition of protein synthesis, elicited by expression of a mutant Ser-51----Asp eIF-2 alpha designed to mimic a phosphorylated serine, is not relieved by the presence of K3L, suggesting that K3L cannot bypass a block imposed by eIF-2 alpha phosphorylation. The results suggest that K3L acts as a decoy of eIF-2 alpha to inhibit DAI kinase autophosphorylation and activation. Another vaccinia virus gene product, K1L, which is required for growth of vaccinia virus on human cells, does not enhance translation in this assay.
Genetics Institute, Cambridge, Massachusetts 02140-2387.
M V
MV
Davies
O
O
Elroy-Stein
R
R
Jagus
B
B
Moss
R J
RJ
Kaufman
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000378:metabolism*; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins / Q000378:metabolism*; D019892:eIF-2 Kinase
D004279:DNA, Viral; D015852:Eukaryotic Initiation Factor-2; D047428:Protein Kinase Inhibitors; D012330:RNA, Double-Stranded; D014764:Viral Proteins; D011494:Protein Kinases; D019892:eIF-2 Kinase