Murine cytomegalovirus m142 and m143 are both required to block protein kinase R-mediated shutdown of protein synthesis.
Cytomegaloviruses carry the US22 family of genes, which have common sequence motifs but diverse functions. Only two of the 12 US22 family genes of murine cytomegalovirus (MCMV) are essential for virus replication, but their functions have remained unknown. In the present study, we deleted the essential US22 family genes, m142 and m143, from the MCMV genome and propagated the mutant viruses on complementing cells. The m142 and the m143 deletion mutants were both unable to replicate in noncomplementing cells at low and high multiplicities of infection. In cells infected with the deletion mutants, viral immediate-early and early proteins were expressed, but viral DNA replication and synthesis of the late-gene product glycoprotein B were inhibited, even though mRNAs of late genes were present. Global protein synthesis was impaired in these cells, which correlated with phosphorylation of the double-stranded RNA-dependent protein kinase R (PKR) and its target protein, the eukaryotic translation initiation factor 2alpha, suggesting that m142 and m143 are necessary to block the PKR-mediated shutdown of protein synthesis. Replication of the m142 and m143 knockout mutants was partially restored by expression of the human cytomegalovirus TRS1 gene, a known double-stranded-RNA-binding protein that inhibits PKR activation. These results indicate that m142 and m143 are both required for inhibition of the PKR-mediated host antiviral response.
Valchanova
Ralitsa S
RS
Robert Koch-Institut, Fachgebiet Virale Infektionen, Nordufer 20, 13353 Berlin, Germany.
Picard-Maureau
Marcus
M
Budt
Matthias
M
Brune
Wolfram
W
D000595:Amino Acid Sequence; D000818:Animals; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D017353:Gene Deletion; D015870:Gene Expression; D020043:Genes, Essential; D005814:Genes, Viral; D005816:Genetic Complementation Test; D051379:Mice; D008969:Molecular Sequence Data; D018146:Muromegalovirus / Q000235:genetics / Q000502:physiology*; D041681:NIH 3T3 Cells; D010766:Phosphorylation; D014176:Protein Biosynthesis; D016415:Sequence Alignment; D014759:Viral Envelope Proteins / Q000096:biosynthesis; D014764:Viral Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014779:Virus Replication / Q000235:genetics; D019892:eIF-2 Kinase / Q000037:antagonists & inhibitors* / Q000378:metabolism
D004279:DNA, Viral; D015852:Eukaryotic Initiation Factor-2; C072216:TRS1 protein, Human herpesvirus 5; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C040495:glycoprotein B, Simplexvirus; D019892:eIF-2 Kinase