Identification of Small Molecule Inhibitors of Pseudomonas aeruginosa Exoenzyme S Using a Yeast Phenotypic Screen

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Identification of Small Molecule Inhibitors of Pseudomonas aeruginosa Exoenzyme S Using a Yeast Phenotypic Screen

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dc.contributor.author Arnoldo, Anthony
dc.contributor.author Curak, Jasna
dc.contributor.author Kittanakom, Saranya
dc.contributor.author Chevelev, Igor
dc.contributor.author Sahebol-Amri, Mehdi
dc.contributor.author Koscik, Becky
dc.contributor.author Ljuma, Lana
dc.contributor.author Roy, Peter J.
dc.contributor.author Bedalov, Antonio
dc.contributor.author Giaever, Guri
dc.contributor.author Nislow, Corey
dc.contributor.author Merrill, Rod A.
dc.contributor.author Stagljar, Igor
dc.contributor.author Snyder, Michael
dc.contributor.author Lee, Vincent
dc.contributor.author Lory, Stephen
dc.date.accessioned 2011-10-31T16:38:35Z
dc.date.issued 2008
dc.identifier.citation Arnoldo, Anthony, Jasna Curak, Saranya Kittanakom, Igor Chevelev, Vincent T. Lee, Mehdi Sahebol-Amri, Becky Koscik, and et al. 2008. Identification of small molecule inhibitors of Pseudomonas aeruginosa exoenzyme S using a yeast phenotypic screen. PLoS Genetics 4(2): e1000005. en_US
dc.identifier.issn 1553-7390 en_US
dc.identifier.uri http://nrs.harvard.edu/urn-3:HUL.InstRepos:5332808
dc.description.abstract Pseudomonas aeruginosa is an opportunistic human pathogen that is a key factor in the mortality of cystic fibrosis patients, and infection represents an increased threat for human health worldwide. Because resistance of Pseudomonas aeruginosa to antibiotics is increasing, new inhibitors of pharmacologically validated targets of this bacterium are needed. Here we demonstrate that a cell-based yeast phenotypic assay, combined with a large-scale inhibitor screen, identified small molecule inhibitors that can suppress the toxicity caused by heterologous expression of selected Pseudomonas aeruginosa ORFs. We identified the first small molecule inhibitor of Exoenzyme S (ExoS), a toxin involved in Type III secretion. We show that this inhibitor, exosin, modulates ExoS ADP-ribosyltransferase activity in vitro, suggesting the inhibition is direct. Moreover, exosin and two of its analogues display a significant protective effect against Pseudomonas infection in vivo. Furthermore, because the assay was performed in yeast, we were able to demonstrate that several yeast homologues of the known human ExoS targets are likely ADP-ribosylated by the toxin. For example, using an in vitro enzymatic assay, we demonstrate that yeast Ras2p is directly modified by ExoS. Lastly, by surveying a collection of yeast deletion mutants, we identified Bmh1p, a yeast homologue of the human FAS, as an ExoS cofactor, revealing that portions of the bacterial toxin mode of action are conserved from yeast to human. Taken together, our integrated cell-based, chemical-genetic approach demonstrates that such screens can augment traditional drug screening approaches and facilitate the discovery of new compounds against a broad range of human pathogens. en_US
dc.language.iso en_US en_US
dc.publisher Public Library of Science en_US
dc.relation.isversionof doi://10.1371/journal.pgen.1000005 en_US
dc.relation.hasversion http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2265467/pdf/ en_US
dash.license LAA
dc.subject biochemistry en_US
dc.subject drug discovery en_US
dc.subject infectious diseases en_US
dc.subject antimicrobials and drug resistance en_US
dc.subject bacterial infections en_US
dc.subject microbiology en_US
dc.subject applied microbiology en_US
dc.title Identification of Small Molecule Inhibitors of Pseudomonas aeruginosa Exoenzyme S Using a Yeast Phenotypic Screen en_US
dc.type Journal Article en_US
dc.description.version Version of Record en_US
dc.relation.journal PLoS Genetics en_US
dash.depositing.author Lory, Stephen
dc.date.available 2011-10-31T16:38:35Z
dash.affiliation.other HMS^Microbiology and Molecular Genetics en_US

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