title: A point mutation in the putative ATP binding site of the pseudorabies virus US3 protein kinase prevents Bad phosphorylation and cell survival following apoptosis induction.
abstract: The multifunctional US3 protein kinase is conserved among alphaherpesviruses. Like the herpes simplex virus US3 protein kinase, the pseudorabies virus (PRV) US3 protein confers resistance against apoptosis. In the current report, we introduced a point mutation in the putative ATP binding site of the PRV US3 protein kinase. We found that, in contrast to the wild type PRV US3, the point-mutated PRV US3 does not protect cells from apoptosis induced by PRV infection or staurosporine treatment. In addition, we found that the presence of wild type PRV US3, but not of the point-mutated PRV US3, results in phosphorylation of the pro-apoptotic Bad protein in PRV-infected ST and HEp-2 cells. In PRV-infected ST cells, but not in HEp-2 cells, an additional, US3- and phosphorylation-independent alteration of Bad could be observed. These results suggest that the kinase activity of the US3 protein of PRV is crucial to protect cells from apoptotic cell death during infection, at least partly by leading to phosphorylation of the pro-apoptotic Bad protein.
authors.affiliation: Department of Virology, Parasitology, and Immunology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium.
authors.forename: Matthias
authors.initials: M
authors.lastname: Deruelle
authors.forename: Kristin
authors.initials: K
authors.lastname: Geenen
authors.forename: Hans J
authors.initials: HJ
authors.lastname: Nauwynck
authors.forename: Herman W
authors.initials: HW
authors.lastname: Favoreel
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis; D001665:Binding Sites / Q000235:genetics; D045744:Cell Line, Tumor / Q000166:cytology; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D006801:Humans; D008297:Male; D010766:Phosphorylation; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics*; D019311:Staurosporine; D013552:Swine; D013737:Testis / Q000166:cytology / Q000821:virology; D014162:Transfection; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics*; D051025:bcl-Associated Death Protein / Q000378:metabolism*
chemical_list: D014764:Viral Proteins; D051025:bcl-Associated Death Protein; D000255:Adenosine Triphosphate; D017346:Protein Serine-Threonine Kinases; D019311:Staurosporine