title: Genome-wide Screen of Pseudomonas aeruginosa in Saccharomyces cerevisiae Identifies New Virulence Factors.
abstract: Pseudomonas aeruginosa is a human opportunistic pathogen that causes mortality in cystic fibrosis and immunocompromised patients. While many virulence factors of this pathogen have already been identified, several remain to be discovered. In this respect we set an unprecedented genome-wide screen of a P. aeruginosa expression library based on a yeast growth phenotype. Fifty-one candidates were selected in athree-round screening process. The robustness of the screen was validated by the selection of three well known secreted proteins including one demonstrated virulence factor, the protease LepA. Further in silico sorting of the 51 candidates highlighted three potential new Pseudomonas effector candidates (Pec). By testing the cytotoxicity of wild type P. aeruginosa vs. pec mutants toward macrophages and the virulence in the Caenorhabditis elegans model, we demonstrated that the three selected Pecs are novel virulence factors of P. aeruginosa. Additional cellular localization experiments in the host revealed specific localization for Pec1 and Pec2 that could inform about their respective functions.
authors.affiliation: Institut für Funktionelle Genomforschung der Mikroorganismen, Heinrich-Heine-Universität Düsseldorf Düsseldorf, Germany.
authors.forename: Rafat
authors.initials: R
authors.lastname: Zrieq
authors.affiliation: Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR7255), Institut de Microbiologie de la Méditerranée, Centre National de la Recherche Scientifique, Aix-Marseille Université Marseille, France.
authors.forename: Thibault G
authors.initials: TG
authors.lastname: Sana
authors.affiliation: Institut für Funktionelle Genomforschung der Mikroorganismen, Heinrich-Heine-Universität Düsseldorf Düsseldorf, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Vergin
authors.affiliation: Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR7255), Institut de Microbiologie de la Méditerranée, Centre National de la Recherche Scientifique, Aix-Marseille Université Marseille, France.
authors.forename: Steve
authors.initials: S
authors.lastname: Garvis
authors.affiliation: Institut für Funktionelle Genomforschung der Mikroorganismen, Heinrich-Heine-Universität Düsseldorf Düsseldorf, Germany.
authors.forename: Irina
authors.initials: I
authors.lastname: Volfson
authors.affiliation: Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR7255), Institut de Microbiologie de la Méditerranée, Centre National de la Recherche Scientifique, Aix-Marseille Université Marseille, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bleves
authors.affiliation: Laboratoire d'Ingénierie des Systèmes Macromoléculaires (UMR7255), Institut de Microbiologie de la Méditerranée, Centre National de la Recherche Scientifique, Aix-Marseille Université Marseille, France.
authors.forename: Romé
authors.initials: R
authors.lastname: Voulhoux
authors.affiliation: Institut für Funktionelle Genomforschung der Mikroorganismen, Heinrich-Heine-Universität Düsseldorf Düsseldorf, Germany.
authors.forename: Johannes H
authors.initials: JH
authors.lastname: Hegemann
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000382:microbiology; D002460:Cell Line; D005820:Genetic Testing / Q000379:methods*; D054884:Host-Pathogen Interactions; D006801:Humans; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000472:pathogenicity*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development*; D037521:Virulence Factors / Q000235:genetics*
chemical_list: D037521:Virulence Factors
keywords: Caenorhabditis elegans model; Pseudomonas aeruginosa; Pseudomonas effector candidates (Pec); genome-wide screening in Saccharomyces cerevisiae; virulence factors