title: Characterization of YopT effects on Rho GTPases in Yersinia enterocolitica-infected cells.
abstract: Pathogenic yersiniae employ a type III secretion system for translocating up to six effector proteins (Yersinia outer proteins (Yops)) into eukaryotic target cells. YopT is a cysteine protease that was shown to remove the C-terminal isoprenoid group of RhoA, Rac, and CDC42Hs. Here we characterized the cell biological and biochemical activities of YopT in cells infected with pathogenic Yersinia enterocolitica. Bacterially injected YopT located to cell membranes from which it released RhoA but not Rac or CDC42Hs. In the infected cells RhoA was dissociated from guanine nucleotide dissociation inhibitor-1 (GDI-1) and accumulated as a monomeric protein in the cytosol, whereas Rac and CDC42Hs remained GDI-bound. Direct transfer of isoprenylated RhoA to YopT and RhoA modification could be reconstituted in vitro by guanosine 5'-3-O-(thio)triphosphate loading of a recombinant RhoA.GDI-1 complex. Finally, in macrophages infected with a Yersinia strain selectively translocating YopT podosomal adhesion structures required for chemotaxis as well as phagocytic cups mediating uptake of yersiniae were disrupted. These findings indicate that bacterially translocated YopT acts on membrane-bound and GDI-complexed RhoA but not Rac or CDC42, and this is sufficient for disruption of macrophage immune functions.
authors.affiliation: Max von Pettenkofer-Institut für Medizinische Mikrobiologie, Ludwig Maximilians-Universität, Pettenkoferstrasse 9a, 80336 Munich, Germany. aepfelbacher@m3401.mpk.med.uni-muenchen.de
authors.forename: Martin
authors.initials: M
authors.lastname: Aepfelbacher
authors.forename: Claudia
authors.initials: C
authors.lastname: Trasak
authors.forename: Gottfried
authors.initials: G
authors.lastname: Wilharm
authors.forename: Agnès
authors.initials: A
authors.lastname: Wiedemann
authors.forename: Konrad
authors.initials: K
authors.lastname: Trulzsch
authors.forename: Kristina
authors.initials: K
authors.lastname: Krauss
authors.forename: Peter
authors.initials: P
authors.lastname: Gierschik
authors.forename: Jurgen
authors.initials: J
authors.lastname: Heesemann
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D015153:Blotting, Western; D002417:Cattle; D002448:Cell Adhesion; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002633:Chemotaxis; D002850:Chromatography, Gel; D003546:Cysteine Endopeptidases; D003600:Cytosol / Q000378:metabolism; D003603:Cytotoxins / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000378:metabolism / Q000382:microbiology; D020730:Guanine Nucleotide Dissociation Inhibitors / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007313:Insecta; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011994:Recombinant Proteins / Q000378:metabolism; D013347:Subcellular Fractions; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology / Q000382:microbiology; D015008:Yersinia enterocolitica / Q000472:pathogenicity*; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D003603:Cytotoxins; D020730:Guanine Nucleotide Dissociation Inhibitors; D011994:Recombinant Proteins; C116117:YopT protein, Yersinia; D003546:Cysteine Endopeptidases; D020764:cdc42 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein