Herpes simplex virus 2 VP22 phosphorylation induced by cellular and viral kinases does not influence intracellular localization.
Phosphorylation of the herpes simplex virus (HSV) VP22 protein is regulated by cellular kinases and the UL13 viral kinase, but the sites at which these enzymes induce phosphorylation of HSV-2 VP22 are not known. Using serine-to-alanine mutants to map phosphorylation sites on HSV-2 VP22 in cells, we made three major observations. First, phosphorylation by a cellular kinase mapped to serines 70, 71, and/or 72 within CKII consensus sites analogous to previously identified phosphorylation sites in HSV-1 VP22. Second, we mapped UL13-mediated phosphorylation of HSV-2 VP22 to serines 28 and 34, describing for the first time UL13-dependent phosphorylation sites on VP22. Third, previously identified VP22-associated cellular kinase sites in HSV-1 VP22 (serines 292 and 294) were not phosphorylated in HSV-2 VP22 (serines 291 and 293). VP22 expressed alone accumulated in the cytoplasm and to a lesser extent in the nucleus. Phosphorylation by endogenous cellular kinase(s) did not alter the localization of VP22. Co-expression of HSV-2 VP22 with active UL13, but not with enzymatically inactive UL13, resulted in nuclear accumulation of VP22 and altered nuclear morphology. Surprisingly, redistribution of VP22 to the nucleus occurred independently of UL13-induced phosphorylation of VP22. The altered nuclear morphology of UL13-expressing cells was not due to apoptosis. These results demonstrate that phosphorylation of HSV-2 VP22 at multiple serine residues is induced by UL13 and cellular kinase(s), and that the nuclear/cytoplasmic distribution of VP22 is independent of its phosphorylation status but is controlled indirectly by UL13 kinase activity.
Geiss
Brian J
BJ
Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, St. Louis, MO 63104-1083, USA.
Cano
Gina L
GL
Tavis
John E
JE
Morrison
Lynda A
LA
D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D018258:Herpesvirus 2, Human / Q000201:enzymology* / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D011494:Protein Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014709:Vero Cells; D015678:Viral Structural Proteins / Q000378:metabolism*
D011994:Recombinant Proteins; D015678:Viral Structural Proteins; C114208:protein VP22, Herpes simplex virus type 2; D011494:Protein Kinases