Serine protein kinase associated with varicella-zoster virus ORF 47.
Varicella-zoster virus (VZV) ORF 47 lies in the unique long region of the VZV genome. Sequence homology studies have demonstrated that gene 47 possessed conserved protein kinase motifs. In this study, we investigated the properties of the ORF 47 product. First, a rabbit antiserum was raised against a protein generated from the fusion of the most antigenic ORF 47 domain with Escherichia coli beta-galactosidase. The high-titer antiserum reacted specifically with ORF 47 polypeptides translated in vitro. When incubated with VZV-infected cell lysate, the antiserum immunoprecipitated a phosphoprotein of M(r) 54,000, a size comparable with the predicted molecular mass. The precipitated viral protein was phosphorylated in a protein kinase assay; subsequent phosphoamino acid analysis indicated that the phosphotransferase associated with the ORF 47 protein was a serine protein kinase. Synthesis of the ORF 47 product in VZV-infected cell culture increased in the first and second days and plateaued after the third day of infection. The protein kinase activity associated with VZV ORF 47 had several distinctive biochemical properties: (i) its phosphotransferase activity was enhanced more by manganese than by magnesium, (ii) it utilized both ATP and GTP as donors of phosphate, and (iii) it phosphorylated both acidic and basic substrates. In summary, this report lends support to the computer homology data which predicted that VZV ORF 47 would encode a serine protein kinase.
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Department of Microbiology, University of Iowa College of Medicine, Iowa City 52242.
Grose
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D000914:Antibodies, Viral / Q000096:biosynthesis; D000918:Antibody Specificity; D002364:Caseins / Q000378:metabolism; D003001:Cloning, Molecular; D005814:Genes, Viral; D006160:Guanosine Triphosphate / Q000378:metabolism; D014645:Herpesvirus 3, Human / Q000201:enzymology / Q000235:genetics*; D006657:Histones / Q000378:metabolism; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics*; D010766:Phosphorylation; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured
D000914:Antibodies, Viral; D002364:Caseins; D006657:Histones; D006160:Guanosine Triphosphate; D017346:Protein Serine-Threonine Kinases