AUTHOR=Zrieq Rafat , Sana Thibault G. , Vergin Sandra , Garvis Steve , Volfson Irina , Bleves Sophie , Voulhoux Romé , Hegemann Johannes H. 

TITLE=Genome-wide Screen of Pseudomonas aeruginosa in Saccharomyces cerevisiae Identifies New Virulence Factors

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=5

YEAR=2015

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2015.00081

DOI=10.3389/fcimb.2015.00081

ISSN=2235-2988

ABSTRACT=<p><italic>Pseudomonas aeruginosa</italic> 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 <italic>P. aeruginosa</italic> 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 <italic>in silico</italic> sorting of the 51 candidates highlighted three potential new <italic>Pseudomonas</italic> effector candidates (Pec). By testing the cytotoxicity of wild type <italic>P. aeruginosa</italic> vs. <italic>pec</italic> mutants toward macrophages and the virulence in the <italic>Caenorhabditis elegans</italic> model, we demonstrated that the three selected Pecs are novel virulence factors of <italic>P. aeruginosa</italic>. Additional cellular localization experiments in the host revealed specific localization for Pec1 and Pec2 that could inform about their respective functions.</p>