title: Phosphorylation of Wzc, a tyrosine autokinase, is essential for assembly of group 1 capsular polysaccharides in Escherichia coli.
abstract: Wzc proteins are tyrosine autokinases. They are found in some important bacterial pathogens of humans and livestock as well as plant-associated bacteria, and are often encoded within gene clusters determining synthesis and assembly of capsular and extracellular polysaccharides. Autophosphorylation of Wzc(cps) is essential for assembly of the serotype K30 group 1 capsule in Escherichia coli O9a:K30, although a genetically unlinked Wzc(cps)-homologue (Etk) can also participate with low efficiency. While autophosphorylation of Wzc(cps) is required for assembly of high molecular weight K30 capsular polysaccharide, it is not essential for either the synthesis of the K30 repeat units or for activity of the K30 polymerase enzyme. Paradoxically, the cognate phosphotyrosine protein phosphatase for Wzc(cps), Wzb(cps), is also required for capsule expression. The tyrosine-rich domain at the C terminus of Wzc(cps) was identified as the site of phosphorylation and autophosphorylation of Wzc requires a functional Walker A motif. Intermolecular transphosphorylation of Wzc(cps) was detected in strains expressing a combination of mutant Wzc(cps) derivatives. The N- and C-terminal domains of Wzc(cps) were expressed independently to mimic the situation found naturally in Gram-positive bacteria. In this format, both domains were required for phosphorylation of the Wzc(cps) C terminus, and for capsule assembly. Regulation by a post-translational phosphorylation event represents a new dimension in the assembly of bacterial cell-surface polysaccharides.
authors.affiliation: Department of Microbiology, University of Guelph, Guelph, Ontario, N1G 2W1 Canada.
authors.forename: T
authors.initials: T
authors.lastname: Wugeditsch
authors.forename: A
authors.initials: A
authors.lastname: Paiment
authors.forename: J
authors.initials: J
authors.lastname: Hocking
authors.forename: J
authors.initials: J
authors.lastname: Drummelsmith
authors.forename: C
authors.initials: C
authors.lastname: Forrester
authors.forename: C
authors.initials: C
authors.lastname: Whitfield
mesh_terms: D000595:Amino Acid Sequence; D000942:Antigens, Bacterial; D000954:Antigens, Surface / Q000096:biosynthesis*; D016667:Bacterial Capsules / Q000096:biosynthesis*; D001426:Bacterial Proteins; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C030659:K antigens; D008565:Membrane Proteins; C417168:WZC protein, Bacteria; D011505:Protein-Tyrosine Kinases; C499615:wzc protein, E coli; D010749:Phosphoprotein Phosphatases; C428435:Wzb protein, E coli