title: Identification of otubain 1 as a novel substrate for the Yersinia protein kinase using chemical genetics and mass spectrometry.
abstract: Yersinia encodes a protein kinase, YpkA, which disrupts the actin cytoskeleton. Using an approach termed chemical genetics, we identified a 36-kDa substrate for YpkA in both J774 lysates and bovine brain cytosol. Mass spectrometry analysis identified this substrate as FLJ20113, an open reading frame that corresponds to otubain 1, a deubiquitinating enzyme implicated in immune cell clonal anergy. We demonstrate that otubain 1 is phosphorylated by YpkA in vitro and interacts with YpkA and actin in vivo. Identification of otubain 1 as a YpkA substrate suggests that regulation of immune cell anergy may be a survival mechanism for Yersinia.
authors.affiliation: Department of Chemistry and Biochemistry, University of Windsor, 401 Sunset Avenue, Windsor, Ont., Canada N9B3P4.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Juris
authors.forename: Kavita
authors.initials: K
authors.lastname: Shah
authors.forename: Kevan
authors.initials: K
authors.lastname: Shokat
authors.forename: Jack E
authors.initials: JE
authors.lastname: Dixon
authors.forename: Panayiotis O
authors.initials: PO
authors.lastname: Vacratsis
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D001923:Brain Chemistry; D002417:Cattle; D002460:Cell Line; D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D017635:Clonal Anergy / Q000187:drug effects / Q000276:immunology; D003546:Cysteine Endopeptidases / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D013379:Substrate Specificity; D015007:Yersinia / Q000201:enzymology* / Q000276:immunology / Q000472:pathogenicity
chemical_list: D001426:Bacterial Proteins; C079893:ypkA protein, Yersinia; D017346:Protein Serine-Threonine Kinases; D003546:Cysteine Endopeptidases; C507756:Otub1 protein, mouse