title: The cyclin-dependent kinase inhibitor roscovitine inhibits the transactivating activity and alters the posttranslational modification of herpes simplex virus type 1 ICP0.
abstract: The cyclin-dependent kinase (cdk) inhibitor Roscovitine (Rosco) reduces transcription of herpes simplex virus early genes significantly, even in the presence of wild-type levels of immediate-early (IE) viral proteins, suggesting that the transactivating functions of IE proteins may require the activities of one or more Rosco-sensitive cdk (L. M. Schang, A. Rosenberg, and P. A. Schaffer, J. Virol. 73:2161-2172, 1999). Based on this observation, we sought to determine whether Rosco alters the transactivating activity and posttranslational modification state of the IE protein, infected cell protein 0 (ICP0), in KOS6beta-infected Vero cells. KOS6beta is a KOS-derived recombinant virus containing an ICP0-inducible ICP6 promoter::lacZ cassette. To monitor ICP0's transactivating activity, KOS6beta-infected cells were released from a cycloheximide (CHX)-mediated protein synthesis block into medium with or without Rosco, and beta-galactosidase activity was measured. Rosco inhibited the ability of ICP0 to transactivate the ICP6 promoter by 50-fold. This inhibition was shown not to be a consequence of inhibition of ICP6 basal promoter activity or aberrant nuclear localization of ICP0. Rosco also altered the electrophoretic mobility of a portion of ICP0 molecules derived from KOS-infected cells following reversal of a CHX block. Notably, however, Rosco had only a minimal effect on the phosphorylation state of ICP0. We conclude that ICP0's transactivating activity requires Rosco-sensitive cdks and hypothesize that these cdks regulate the functions of cellular enzymes which modify ICP0, and are, consequently, required for its transactivating activity. Thus, we propose that Rosco regulates ICP0's posttranslational state by mechanisms other than, or in addition to, phosphorylation.
authors.affiliation: Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6076, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Davido
authors.forename: David A
authors.initials: DA
authors.lastname: Leib
authors.forename: Priscilla A
authors.initials: PA
authors.lastname: Schaffer
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015870:Gene Expression; D005818:Genetic Engineering; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D017874:Immediate-Early Proteins / Q000187:drug effects* / Q000378:metabolism; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000032:analysis; D010766:Phosphorylation / Q000187:drug effects; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011687:Purines / Q000494:pharmacology*; D000077546:Roscovitine; D015534:Trans-Activators / Q000187:drug effects* / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects*; D014357:Trypsin; D044767:Ubiquitin-Protein Ligases; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D004791:Enzyme Inhibitors; D017874:Immediate-Early Proteins; D010750:Phosphoproteins; D011687:Purines; D015534:Trans-Activators; D014764:Viral Proteins; C054366:herpes simplex virus type 1-protein ICP6; D000077546:Roscovitine; D044767:Ubiquitin-Protein Ligases; C051804:Vmw110 protein, Human herpesvirus 1; D018844:Cyclin-Dependent Kinases; D001616:beta-Galactosidase; D014357:Trypsin