Isolation of a protein kinase induced by herpes simplex virus type 1

Varicella-zoster virus (VZV) glycoprotein gpl is the predominant viral glycoprotein within the plasma membranes of infected cells. This viral glycoprotein is phosphorylated on its polypeptide backbone during biosynthesis. In this report, we investigated the protein kinases which participate in the phosphorylation events. Under in vivo conditions, VZV gpl was phosphorylated on its serine and threonine residues by protein kinases present within lysates of either VZV-infected or uninfected cells. Because this activity was diminished by heparin, a known inhibitor of casein kinase II, isolated gpl was incubated with purified casein kinase II and shown to be phosphorylated in an in vitro assay containing [y-32P]ATP. The same glycoprotein was phosphorylated when [32P]GTP was substituted for [32P]ATP in the protein kinase assay. We also tested whether VZV gpl was phosphorylated by two other ubiquitous mammalian protein kinases-casein kinase I and cyclic AMP-dependent kinase-and found that only casein kinase I modified gpl. When the predicted 623-amino-acid sequence of gpl was examined, two phosphorylation sites known to be optimal for casein kinase II were observed. Immediately upstream from each of the casein kinase II sites was a potential casein kinase I phosphorylation site. In summary, this study showed that VZV gpl was phosphorylated by each of two mammalian protein kinases (casein kinase I and casein kinase II) and that potential serine-threonine phosphorylation sites for each of these two kinases were present in the viral glycoprotein.

supporting

confidence: 55%

Phosphorylation of varicella-zoster virus glycoprotein gpI by mammalian casein kinase II and casein kinase I

Grose

Jackson

Traugh

1989

“…Biochem., in press) and highly purified from cells infected with HSV-1 (27). The substrate specificity and other properties of the enzyme clearly differentiate it from known c'ellular protein kinases (13) and from another activity reported in cells infected with HSV-1 and catalyzing the phosphorylation of the acidic protein phosvitin in vitro (3). Furthermore, the protein kinases induced in cells infected with HSV-1 and PRV could be chromatographically resolved from one another, suggesting, although not proving, that they are viral gene products.…”

mentioning

confidence: 73%

Herpes simplex virus 1 protein kinase is encoded by open reading frame US3 which is not essential for virus growth in cell culture

Purves¹,

Longnecker²,

Leader³

et al. 1987

181

Earlier reports have described a novel protein kinase in cells infected with herpes simplex or pseudorabies viruses. These novel enzymes were characterized by their acceptance of protamine as a substrate and by their differential chromatographic behavior in anion-exchange chromatography. We report that this activity was not present in extracts of uninfected cells or of cells infected with a mutant constructed so as to contain a deletion in the US3 open reading frame mapping in the small component of herpes simplex virus 1 DNA. The activity was present in extracts of cells infected with wild-type virus and with a recombinant in which the US3 open reading frame had been rescued. Our results are consistent with the observation reported earlier that the coding sequences predict an amino acid motif common to protein kinases and lead to the conclusion that the US3 open reading frame encodes-a virus-specific protein kinase that is not required for virus growth in cells in culture.

“…A number of workers have proposed that RNA polymerases are partially regulated by a phosphorylationdephosphorylation mechanism (7,8,20,21,37,40). Phosphorylation of proteins also appears to play a role in the control of HSV infections (6,22,30,50). There is a protein kinase activity associated with purified virions (22, 38) and an HSV-specific protein kinase activity which appears within the first hour of infection (6).…”

Section: D)iscussionmentioning

confidence: 99%

Herpes simplex virus-induced changes in cellular and adenovirus RNA metabolism in an adenovirus type 5-transformed human cell line

Stenberg¹,

Pizer²

1982

We used the viral transcripts (designated Ad-RNA) that accumulated in the cytoplasm of adenovirus type 5-transformed human embryonic kidney cells (cell line 293-31) as models for cellular RNAs to examine how herpes simplex virus modifies cellular RNA metabolism. Infection of 293-31 cells with herpes simplex virus type 1 strain 17 led to extensive inhibition of Ad-RNA accumulation by 4 h postinfection. The major part of this inhibition was due to an immediate early or alpha gene function, which reduced the rate of transcription of Ad-RNA within the nuclei of the infected cells. In addition, host polyadenylic acid-containing RNA accumulation and rRNA accumulation were affected, but to a lesser extent and at a slower rate than Ad-RNA accumulation. In conjunction with previous data, our experimental data allowed us to propose a general scheme for how herpes simplex virus type 1 alters the metabolism of cellular RNA, the possible mechanisms for these changes, and how they correlate with the regulation of herpes simplex virus gene expression.