Abstract
Varicella-zoster virus (VZV) codes for a protein serine kinase called ORF47; the herpes simplex virus (HSV) homolog is UL13. No recombinant alphaherpesvirus serine kinase has been biologically active in vitro. We discovered that preservation of the intrinsic kinase activity of recombinant VZV ORF47 required unusually stringent in vitro conditions, including physiological concentrations of polyamines. In this assay, ORF47 phosphorylated two VZV regulatory proteins: the ORF62 protein (homolog of HSV ICP4) and the ORF63 protein (homolog of HSV ICP22). Of interest, ORF47 kinase also coprecipitated ORF63 protein from the kinase assay supernatant.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Aspartic Acid / genetics
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Aspartic Acid / metabolism
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Baculoviridae
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Cloning, Molecular
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Enzyme Activation
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Genetic Vectors
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HeLa Cells
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Herpesvirus 3, Human / enzymology*
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Humans
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Immediate-Early Proteins / genetics
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Immediate-Early Proteins / metabolism*
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Lysine / genetics
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Lysine / metabolism
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Mutagenesis, Site-Directed
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Phosphorylation
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Polyamines / pharmacology
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Polymerase Chain Reaction / methods
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Protein Kinases / genetics
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Protein Kinases / metabolism*
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Substrate Specificity
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Trans-Activators / genetics
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Trans-Activators / metabolism*
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Viral Envelope Proteins / genetics
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Viral Envelope Proteins / metabolism*
Substances
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IE62 protein, Human herpesvirus 3
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Immediate-Early Proteins
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Polyamines
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Recombinant Fusion Proteins
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Trans-Activators
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Viral Envelope Proteins
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immediate early protein 63, Human herpesvirus 3
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Aspartic Acid
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Protein Kinases
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Varicella-Zoster ORF47-associated protein kinase
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Protein Serine-Threonine Kinases
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Lysine