title: Characterization of a eukaryotic-like tyrosine protein kinase expressed by the Shiga toxin-encoding bacteriophage 933W.
abstract: The Shiga toxin (Stx)-encoding bacteriophage 933W contains an open reading frame, stk, with amino acid sequence similarity to the catalytic domain of eukaryotic serine/threonine (Ser/Thr) protein kinases (PKs). Eukaryotic PKs are related by a common catalytic domain, consisting of invariant and nearly invariant residues necessary for ATP binding and phosphotransfer. We demonstrate that rather than a Ser/Thr kinase, stk encodes a eukaryotic-like tyrosine (Tyr) kinase. An affinity-purified recombinant Stk (rStk) autophosphorylates and catalyzes the phosphorylation of an artificial substrate on Tyr residues and not on Ser or Thr residues. A change of an invariant lysine within the putative catalytic domain abolishes this kinase activity, indicating that Stk uses a phosphotransfer mechanism similar to the mechanism used by eukaryotic PKs. We provide evidence suggesting that stk is cotranscribed with cI from the phage promoter responsible for maintaining CI expression during lysogeny. The stk gene was identified in prophages obtained from independently isolated Stx-producing Escherichia coli clinical isolates, suggesting that selective pressure has maintained the stk gene in these pathogenic bacteria.
authors.affiliation: Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan 48109-0620, USA.
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Tyler
authors.forename: David I
authors.initials: DI
authors.lastname: Friedman
mesh_terms: D000595:Amino Acid Sequence; D003090:Coliphages / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics*; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000235:genetics*; D022621:Shiga Toxin / Q000235:genetics*
chemical_list: D022621:Shiga Toxin; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases