title: Sites and roles of phosphorylation of the human cytomegalovirus DNA polymerase subunit UL44.
abstract: The human cytomegalovirus DNA polymerase subunit UL44 is a phosphoprotein, but its sites and roles of phosphorylation have not been investigated. We compared sites of phosphorylation of UL44 in vitro by the viral protein kinase UL97 and cyclin-dependent kinase 1 with those in infected cells. Transient treatment of infected cells with a UL97 inhibitor greatly reduced labeling of two minor UL44 phosphopeptides. Viruses containing alanine substitutions of most UL44 residues that are phosphorylated in infected cells exhibited at most modest effects on viral DNA synthesis and yield. However, substitution of highly phosphorylated sites adjacent to the nuclear localization signal abolished viral replication. The results taken together are consistent with UL44 being phosphorylated directly by UL97 during infection, and a crucial role for phosphorylation-mediated nuclear localization of UL44 for viral replication, but lend little support to the widely held hypothesis that UL97-mediated phosphorylation of UL44 is crucial for viral DNA synthesis.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Laurie A
authors.initials: LA
authors.lastname: Silva
authors.forename: Blair L
authors.initials: BL
authors.lastname: Strang
authors.forename: Eric W
authors.initials: EW
authors.lastname: Lin
authors.forename: Jeremy P
authors.initials: JP
authors.lastname: Kamil
authors.forename: Donald M
authors.initials: DM
authors.lastname: Coen
mesh_terms: D021581:Active Transport, Cell Nucleus; D019943:Amino Acid Substitution / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism*; D003587:Cytomegalovirus / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D004268:DNA-Binding Proteins; C072217:ICP36 protein, Cytomegalovirus; D014764:Viral Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase