Eukaryotic elongation factor 1delta is hyperphosphorylated by the protein kinase encoded by the U(L)13 gene of herpes simplex virus 1.
The translation elongation factor 1delta (EF-1delta) consists of two forms, a hypophosphorylated form (apparent Mr, 38,000) and a hyperphosphorylated form (apparent Mr, 40,000). Earlier Y. Kawaguchi, R. Bruni, and B. Roizman (J. Virol. 71:1019-1024, 1997) reported that whereas mock-infected cells accumulate the hypophosphorylated form, the hyperphosphorylated form of EF-1delta accumulates in cells infected with herpes simplex virus 1. We now report that the accumulation of the hyperphosphorylated EF-1delta is due to phosphorylation by U(L)13 protein kinase based on the following observations. (i) The relative amounts of hypo- and hyperphosphorylated EF-1delta in Vero cells infected with mutant virus lacking the U(L)13 gene could not be differentiated from those of mock-infected cells. In contrast, the hyperphosphorylated EF-1delta was the predominant form in Vero cells infected with wild-type viruses, a recombinant virus in which the deleted U(L)13 sequences were restored, or with a virus lacking the U(S)3 gene, which also encodes a protein kinase. (ii) The absence of the hyperphosphorylated EF-1delta in cells infected with the U(L)13 deletion mutant was not due to failure of posttranslational modification of infected-cell protein 22 (ICP22)/U(S)1.5 or of interaction with ICP0, inasmuch as preferential accumulation of hyperphosphorylated EF-1delta was observed in cells infected with viruses from which the genes encoding ICP22/U(S)1.5 or ICP0 had been deleted. (iii) Both forms of EF-1delta were labeled by 32Pi in vivo, but the prevalence of the hyperphosphorylated EF-1delta was dependent on the presence of the U(L)13 protein. (iv) EF-1delta immunoprecipitated from uninfected Vero cells was phosphorylated by U(L)13 precipitated by the anti-U(L)13 antibody from lysates of wild-type virus-infected cells, but not by complexes formed by the interaction of the U(L)13 antibody with lysates of cells infected with a mutant lacking the U(L)13 gene. This is the first evidence that a viral protein kinase targets a cellular protein. Together with evidence that ICP0 also interacts with EF-1delta reported in the paper cited above, these data indicate that herpes simplex virus 1 has evolved a complex strategy for optimization of infected-cell protein synthesis.
Kawaguchi
Y
Y
The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, Illinois 60637, USA.
Van Sant
C
C
Roizman
B
B
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D005057:Eukaryotic Cells; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000378:metabolism; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000378:metabolism*; D010766:Phosphorylation; D011233:Precipitin Tests; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D044767:Ubiquitin-Protein Ligases; D014709:Vero Cells; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
C057294:ICP22 protein, human herpesvirus 1; D017874:Immediate-Early Proteins; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C078424:EUS1 protein, Equine herpesvirus 1; D044767:Ubiquitin-Protein Ligases; C051804:Vmw110 protein, Human herpesvirus 1; D011494:Protein Kinases; C082380:UL13 protein, Simplexvirus; D017346:Protein Serine-Threonine Kinases; C095207:US3 protein, Human herpesvirus 1