Genome-Wide Fitness Test and Mechanism-of-Action Studies of Inhibitory Compounds in Candida albicans

PLoS Pathogens

Nick Martel¹  Steve Trosok¹  Bo Jiang¹  John Davison¹  Terry Roemer¹  Deming Xu¹  Susan Sillaots¹  Karynn Veillette¹  Catherine Bachewich²  Howard Bussey³  Phil Youngman⁴  Troy Ketela⁵  Sebastien Lemieux⁶ 

[1] Center of Fungal Genetics, Merck Frosst Canada Ltd., Montreal, Quebec, Canada;Department of Biology, Concordia University, Montreal, Quebec, Canada;Department of Biology, McGill University, Montreal, Quebec, Canada;Department of Infectious Disease, Merck & Co., Inc., Rahway, New Jersey, United States of America;Infinity Pharmaceuticals, Cambridge, Massachusetts, United States of America;Institute of Research in Immunology and Cancer, University of Montreal, Montreal, Quebec, Canada

关键词: C;    ida albicans;    Saccharomyces cerevisiae;    Hypersensitivity;    Antifungals;    Microtubules;    Antimicrobial resistance;    Ribosomal RNA;    Gene pool;   

DOI  :  10.1371/journal.ppat.0030092

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Candida albicans is a prevalent fungal pathogen amongst the immunocompromised population, causing both superficial and life-threatening infections. Since C. albicans is diploid, classical transmission genetics can not be performed to study specific aspects of its biology and pathogenesis. Here, we exploit the diploid status of C. albicans by constructing a library of 2,868 heterozygous deletion mutants and screening this collection using 35 known or novel compounds to survey chemically induced haploinsufficiency in the pathogen. In this reverse genetic assay termed the fitness test, genes related to the mechanism of action of the probe compounds are clearly identified, supporting their functional roles and genetic interactions. In this report, chemical–genetic relationships are provided for multiple FDA-approved antifungal drugs (fluconazole, voriconazole, caspofungin, 5-fluorocytosine, and amphotericin B) as well as additional compounds targeting ergosterol, fatty acid and sphingolipid biosynthesis, microtubules, actin, secretion, rRNA processing, translation, glycosylation, and protein folding mechanisms. We also demonstrate how chemically induced haploinsufficiency profiles can be used to identify the mechanism of action of novel antifungal agents, thereby illustrating the potential utility of this approach to antifungal drug discovery.

【 授权许可】

CC BY