Temporal regulation of herpes simplex virus type 2 VP22 expression and phosphorylation.
The VP22 protein of herpes simplex virus type 2 (HSV-2) is a major component of the virion tegument. Previous work with HSV-1 indicated that VP22 is phosphorylated during infection, and phosphorylation may play a role in modulating VP22 localization in infected cells. It is not clear, however, when phosphorylation occurs in infected cells or how it is regulated. Less is known about the synthesis and phosphorylation of HSV-2 VP22. To study the complete biosynthetic history of HSV-2 VP22, we generated a monoclonal antibody to the carboxy terminus of VP22. Using immunoprecipitation and Western blot analyses, we show that HSV-2 VP22 can be found in three distinct isoforms in infected cells, two of which are phosphorylated. Like HSV-1 VP22, HSV-2 VP22 is synthesized ca. 4 h after infection, and the isoform later incorporated into virions is hypophosphorylated. In addition, we demonstrate for the first time (i) that newly synthesized VP22 is phosphorylated rapidly after synthesis, (ii) that this phosphorylation occurs in a virus-dependent manner, (iii) that the HSV-2 kinase UL13 is capable of inducing phosphorylation of VP22 in the absence of other viral proteins, (iv) that phosphorylated VP22 is very stable in infected cells, (v) that phosphorylated isoforms of VP22 are gradually dephosphorylated late in infection to produce the virion tegument form, and (vi) that this dephosphorylation occurs independently of viral DNA replication or virion assembly. These results indicate that HSV-2 VP22 is a stable protein that undergoes highly regulated, virus-dependent phosphorylation events in infected cells.
Geiss
B J
BJ
Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, St. Louis, Missouri 63104, USA.
Tavis
J E
JE
Metzger
L M
LM
Leib
D A
DA
Morrison
L A
LA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002522:Chlorocebus aethiops; D018258:Herpesvirus 2, Human / Q000378:metabolism* / Q000472:pathogenicity; D010766:Phosphorylation; D020033:Protein Isoforms; D011494:Protein Kinases / Q000502:physiology; D014162:Transfection; D014709:Vero Cells; D015678:Viral Structural Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D020033:Protein Isoforms; D015678:Viral Structural Proteins; C114208:protein VP22, Herpes simplex virus type 2; D011494:Protein Kinases; C082380:UL13 protein, Simplexvirus