UL34, the target of the herpes simplex virus U(S)3 protein kinase, is a membrane protein which in its unphosphorylated state associates with novel phosphoproteins.
Previous studies (F. C. Purves, D. Spector, and B. Roizman, J. Virol. 65:5757-5764, 1991) have shown that the protein kinase encoded by the U(S)3 gene mediates posttranslational modification of a viral phosphoprotein with an apparent M(r) of 30,000 encoded by the UL34 gene. Here we report the following. (i) UL34 protein is not phosphorylated in cells infected with recombinant viruses deleted in the U(S)3 gene. (ii) Several new phosphoproteins (apparent M(r)s, 25,000 to 35,000) are present in cells infected with recombinant viruses deleted in the U(S)3 gene or with viruses carrying a mutation in the UL34 gene that precluded phosphorylation of the UL34 gene product by the U(S)3 protein kinase, but not in cells infected under conditions which permit phosphorylation of the UL34 protein. These proteins are genetically unrelated to the product of the UL34 gene. (iii) Polyclonal rabbit anti-UL34 protein serum precipitated not only the UL34 protein but also the other (25,000- to 35,000-M(r)) phosphoproteins from lysates of cells infected with U(S)3- virus. (iv) The UL34 gene product is a membrane protein inasmuch as the polyclonal anti-UL34 serum reacted with surfaces of intact, unfixed, infected cells and the antigen-antibody complex formed in this reaction contained the UL34 protein. (v) Small amounts of the UL34 protein were present in virions of infected cells. We conclude that the UL34 gene product is a membrane protein exclusively phosphorylated by the U(S)3 protein kinase which can either directly or indirectly form complexes with several other phosphoproteins. Experiments done thus far suggest that these phosphoproteins are present only under conditions in which the UL34 protein is not phosphorylated.
Purves
F C
FC
Marjorie B. Kovler Viral Oncology Laboratory, University of Chicago, Illinois 60637.
Spector
D
D
Roizman
B
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004279:DNA, Viral; D015151:Immunoblotting; D008320:Maltose / Q000378:metabolism; D058619:Maltose-Binding Proteins; D008565:Membrane Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018139:Simplexvirus / Q000201:enzymology*; D013379:Substrate Specificity; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014771:Virion / Q000378:metabolism
D002352:Carrier Proteins; D004279:DNA, Viral; D058619:Maltose-Binding Proteins; D008565:Membrane Proteins; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D008320:Maltose; D011494:Protein Kinases