title: Tyrosine glycosylation of Rho by Yersinia toxin impairs blastomere cell behaviour in zebrafish embryos.
abstract: Yersinia species cause zoonotic infections, including enterocolitis and plague. Here we studied Yersinia ruckeri antifeeding prophage 18 (Afp18), the toxin component of the phage tail-derived protein translocation system Afp, which causes enteric redmouth disease in salmonid fish species. Here we show that microinjection of the glycosyltransferase domain Afp18(G) into zebrafish embryos blocks cytokinesis, actin-dependent motility and cell blebbing, eventually abrogating gastrulation. In zebrafish ZF4 cells, Afp18(G) depolymerizes actin stress fibres by mono-O-GlcNAcylation of RhoA at tyrosine-34; thereby Afp18(G) inhibits RhoA activation by guanine nucleotide exchange factors, and blocks RhoA, but not Rac and Cdc42 downstream signalling. The crystal structure of tyrosine-GlcNAcylated RhoA reveals an open conformation of the effector loop distinct from recently described structures of GDP- or GTP-bound RhoA. Unravelling of the molecular mechanism of the toxin component Afp18 as glycosyltransferase opens new perspectives in studies of phage tail-derived protein translocation systems, which are preserved from archaea to human pathogenic prokaryotes.
authors.affiliation: Institute of Experimental and Clinical Pharmacology and Toxicology, Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Jank
authors.affiliation: Department of Developmental Biology, Institute Biology I, Faculty of Biology, Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Eckerle
authors.affiliation: Institute of Experimental and Clinical Pharmacology and Toxicology, Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Steinemann
authors.affiliation: Institute of Experimental and Clinical Pharmacology and Toxicology, Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Trillhaase
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
authors.forename: Marianne
authors.initials: M
authors.lastname: Schimpl
authors.affiliation: Center for Biological Systems Analysis, Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Wiese
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, UK.
authors.forename: Daan M F
authors.initials: DM
authors.lastname: van Aalten
authors.affiliation: 1] Department of Developmental Biology, Institute Biology I, Faculty of Biology, Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany [2] Centre for Biological Signalling Studies (BIOSS), Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Driever
authors.affiliation: 1] Institute of Experimental and Clinical Pharmacology and Toxicology, Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany [2] Centre for Biological Signalling Studies (BIOSS), Albert-Ludwigs-University Freiburg, D-79104 Freiburg, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Aktories
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology*; D001757:Blastomeres / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D048749:Cytokinesis / Q000187:drug effects*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D006031:Glycosylation; D016695:Glycosyltransferases / Q000494:pharmacology*; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D020559:Monomeric GTP-Binding Proteins / Q000187:drug effects* / Q000378:metabolism; D011487:Protein Conformation / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D014443:Tyrosine / Q000187:drug effects* / Q000378:metabolism; D044124:Yersinia ruckeri; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000187:drug effects* / Q000378:metabolism
chemical_list: D001427:Bacterial Toxins; D020662:Guanine Nucleotide Exchange Factors; D029961:Zebrafish Proteins; D014443:Tyrosine; D016695:Glycosyltransferases; D020559:Monomeric GTP-Binding Proteins; C549425:rhoab protein, zebrafish