title: The Escherichia coli effector EspJ blocks Src kinase activity via amidation and ADP ribosylation.
abstract: The hallmark of enteropathogenic Escherichia coli (EPEC) infection is the formation of actin-rich pedestal-like structures, which are generated following phosphorylation of the bacterial effector Tir by cellular Src and Abl family tyrosine kinases. This leads to recruitment of the Nck-WIP-N-WASP complex that triggers Arp2/3-dependent actin polymerization in the host cell. The same phosphorylation-mediated signalling network is also assembled downstream of the Vaccinia virus protein A36 and the phagocytic Fc-gamma receptor FcγRIIa. Here we report that the EPEC type-III secretion system effector EspJ inhibits autophosphorylation of Src and phosphorylation of the Src substrates Tir and FcγRIIa. Consistent with this, EspJ inhibits actin polymerization downstream of EPEC, Vaccinia virus and opsonized red blood cells. We identify EspJ as a unique adenosine diphosphate (ADP) ribosyltransferase that directly inhibits Src kinase by simultaneous amidation and ADP ribosylation of the conserved kinase-domain residue, Src E310, resulting in glutamine-ADP ribose.
authors.affiliation: MRC Centre for Molecular Bacteriology and Infection, Imperial College, SW7 2AZ London, UK.
authors.forename: Joanna C
authors.initials: JC
authors.lastname: Young
authors.affiliation: MRC Centre for Molecular Bacteriology and Infection, Imperial College, SW7 2AZ London, UK.
authors.forename: Abigail
authors.initials: A
authors.lastname: Clements
authors.affiliation: Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, D-79104 Freiburg, Germany.
authors.forename: Alexander E
authors.initials: AE
authors.lastname: Lang
authors.affiliation: Centre for Structural Biology, Imperial College, SW7 2AZ London, UK.
authors.forename: James A
authors.initials: JA
authors.lastname: Garnett
authors.affiliation: MRC Centre for Molecular Bacteriology and Infection, Imperial College, SW7 2AZ London, UK.
authors.forename: Diana
authors.initials: D
authors.lastname: Munera
authors.affiliation: MRC Centre for Molecular Bacteriology and Infection, Imperial College, SW7 2AZ London, UK.
authors.forename: Ana
authors.initials: A
authors.lastname: Arbeloa
authors.affiliation: Department of Microbiology and Immunology, University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Melbourne Victoria 3010, Australia.
authors.forename: Jaclyn
authors.initials: J
authors.lastname: Pearson
authors.affiliation: Department of Microbiology and Immunology, University of Melbourne, at the Peter Doherty Institute for Infection and Immunity, Melbourne Victoria 3010, Australia.
authors.forename: Elizabeth L
authors.initials: EL
authors.lastname: Hartland
authors.affiliation: Centre for Structural Biology, Imperial College, SW7 2AZ London, UK.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Matthews
authors.affiliation: MRC Centre for Molecular Bacteriology and Infection, Imperial College, SW7 2AZ London, UK.
authors.forename: Aurelie
authors.initials: A
authors.lastname: Mousnier
authors.affiliation: Cell Motility Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Barry
authors.affiliation: Cell Motility Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Way
authors.affiliation: Rudolf Virchow Center for Experimental Biomedicine, University of Wuerzburg, 97080 Würzburg, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Schlosser
authors.affiliation: 1] Institute of Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, D-79104 Freiburg, Germany [2] Centre for Biological Signalling Studies (BIOSS), University of Freiburg, D-79104 Freiburg, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Aktories
authors.affiliation: MRC Centre for Molecular Bacteriology and Infection, Imperial College, SW7 2AZ London, UK.
authors.forename: Gad
authors.initials: G
authors.lastname: Frankel
mesh_terms: D000244:Adenosine Diphosphate / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs; D054324:Enterohemorrhagic Escherichia coli / Q000235:genetics / Q000378:metabolism*; D004927:Escherichia coli Infections / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D017452:Receptors, IgG / Q000378:metabolism; D019061:src-Family Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; C497366:EspJ protein, E coli; C080568:Fc gamma receptor IIA; D017452:Receptors, IgG; D000244:Adenosine Diphosphate; D019061:src-Family Kinases