title: Inhibition of protein kinases C prevents murine cytomegalovirus replication.
abstract: For successful establishment of infection and initiation of the replication cycle, murine cytomegalovirus (MCMV) utilizes cellular structures and functions, including cell-membrane penetration, capsid dismantling and cytosolic transport of viral DNA into the nucleus. These early events of MCMV infections are dependent on cellular regulatory mechanisms, primarily protein phosphorylation. In the present study, protein kinase inhibitors were used to explore the role of protein phosphorylation mediated by protein kinases C (PKCs) in the very early events of MCMV infection. Inhibitory effects were determined by immunofluorescence and Western blot analysis of MCMV IE1 and E1 protein expression and by production of infectious virions in cell culture. It was found that H-7, a broadly specific inhibitor of cellular protein kinases, prevented virus replication in a dose-dependent and reversible manner, and that the block in replication occurred very early in infection. More specific PKC inhibitors (sangivamycin, calphostin C and bisindolylmaleimide II), Ca(2+)/calmodulin inhibitors (EDTA and W7) and phorbol esters (PMA) were used to dissect PKC-subclass contribution in the very early events of MCMV replication. The results indicate that the role of diacylglycerol/phorbol ester-dependent but calcium-independent PKCs is essential for establishment of MCMV infection in the host cell, starting at a very early stage of infection.
authors: Department of Physiology and Immunology, Medical Faculty, University of Rijeka, Braće Branchetta 20, 51000 Rijeka, Croatia.
authors: Natalia
authors: N
authors: Kučić
authors: Department of Physiology and Immunology, Medical Faculty, University of Rijeka, Braće Branchetta 20, 51000 Rijeka, Croatia.
authors: Hana
authors: H
authors: Mahmutefendić
authors: Department of Physiology and Immunology, Medical Faculty, University of Rijeka, Braće Branchetta 20, 51000 Rijeka, Croatia.
authors: Pero
authors: P
authors: Lučin
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006566:Herpesviridae Infections / Q000821:virology*; D051379:Mice; D018146:Muromegalovirus / Q000378:metabolism / Q000502:physiology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism / Q000502:physiology*; D014779:Virus Replication / Q000187:drug effects* / Q000502:physiology
chemical_list: D004791:Enzyme Inhibitors; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C