Viral host-range factor C7 or K1 is essential for modified vaccinia virus Ankara late gene expression in human and murine cells, irrespective of their capacity to inhibit protein kinase R-mediated phosphorylation of eukaryotic translation initiation factor 2alpha.
Vaccinia virus (VACV) infection induces phosphorylation of eukaryotic translation initiation factor 2alpha (eIF2alpha), which inhibits cellular and viral protein synthesis. In turn, VACV has evolved the capacity to antagonize this antiviral response by expressing the viral host-range proteins K3 and E3. This study revealed that the host-range genes K1L and C7L also prevent eIF2alpha phosphorylation in modified VACV Ankara (MVA) infection of several human and murine cell lines. Moreover, C7L-deleted MVA (MVA-DeltaC7L) lacked late gene expression, which could be rescued by the function of host-range factor K1 or C7. It was demonstrated that viral gene expression was blocked after viral DNA replication and that it was independent of apoptosis induction. Furthermore, it was found that eIF2alpha phosphorylation in MVA-DeltaC7L-infected cells is mediated by protein kinase R (PKR) as shown in murine embryonic fibroblasts lacking PKR function, and it was shown that this was not due to reduced E3L gene expression. The block of eIF2alpha phosphorylation by C7 could be complemented by K1 in cells infected with MVA-DeltaC7L encoding a reinserted K1L gene (MVA-DeltaC7L-K1L). Importantly, these data illustrated that eIF2alpha phosphorylation by PKR is not responsible for the block of late viral gene expression. This suggests that other mechanisms targeted by C7 and K1 are essential for completing the MVA gene expression cycle and probably also for VACV replication in a diverse set of cell types.
Institute of Virology, Technical University of Munich, and Helmholtz Center Munich, Germany.
Simone
S
Backes
Karin M
KM
Sperling
Joachim
J
Zwilling
Georg
G
Gasteiger
Holger
H
Ludwig
Elisabeth
E
Kremmer
Astrid
A
Schwantes
Caroline
C
Staib
Gerd
G
Sutter
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D006801:Humans; D051379:Mice; D010766:Phosphorylation; D014615:Vaccinia / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D019892:eIF-2 Kinase / Q000235:genetics / Q000378:metabolism*
C082283:C7L protein, Vaccinia virus; D015852:Eukaryotic Initiation Factor-2; C083205:K1L protein, Vaccinia virus; D014764:Viral Proteins; D019892:eIF-2 Kinase