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Genome-wide screen of Pseudomonas aeruginosa In Saccharomyces cerevisiae identifies new virulence factors

By Rafat eZrieq, Rafat eZrieq, Thibault G Sana, Sandra eVergin, Steve eGarvis, Irina eVolfson, Sophie eBleves, Romé eVoulhoux and Johannes H Hegemann


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. 51 candidates were selected in a three-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 towards 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

Topics: Pseudomonas aeruginosa, Virulence Factors, genome-wide screening in Saccharomyces cerevisiae, Caenorhabditis elegans model, Pseudomonas effector candidates (Pec), Microbiology, QR1-502
Publisher: Frontiers Media S.A.
Year: 2015
DOI identifier: 10.3389/fcimb.2015.00081
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