Essential Gene Identification and Drug Target Prioritization in Aspergillus fumigatus

PLoS Pathogens

Jeff Becker¹  Sarah Kauffman¹  Joel Bowman²  Susan Sillaots³  Chunlin Xin³  Sebastien Lemieux³  Bo Jiang³  John Davison³  Terry Roemer³  Wenqi Hu³  Annie Linteau³  Anouk Breton³ 

[1] Department of Microbiology, University of Tennessee, Knoxville, Tennessee, United States of America;Infectious Diseases, Merck Research Laboratories, Rahway, New Jersey, United States of America;Merck Frosst Center of Fungal Genetics, Montreal, Quebec, Canada

关键词: Aspergillus fumigatus;    Phenotypes;    Mouse models;    Animal models of infection;    Polymerase chain reaction;    C;    ida albicans;    Saccharomyces cerevisiae;    Antifungals;   

DOI  :  10.1371/journal.ppat.0030024

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Aspergillus fumigatus is the most prevalent airborne filamentous fungal pathogen in humans, causing severe and often fatal invasive infections in immunocompromised patients. Currently available antifungal drugs to treat invasive aspergillosis have limited modes of action, and few are safe and effective. To identify and prioritize antifungal drug targets, we have developed a conditional promoter replacement (CPR) strategy using the nitrogen-regulated A. fumigatus NiiA promoter (pNiiA). The gene essentiality for 35 A. fumigatus genes was directly demonstrated by this pNiiA-CPR strategy from a set of 54 genes representing broad biological functions whose orthologs are confirmed to be essential for growth in Candida albicans and Saccharomyces cerevisiae. Extending this approach, we show that the ERG11 gene family (ERG11A and ERG11B) is essential in A. fumigatus despite neither member being essential individually. In addition, we demonstrate the pNiiA-CPR strategy is suitable for in vivo phenotypic analyses, as a number of conditional mutants, including an ERG11 double mutant (erg11BΔ, pNiiA-ERG11A), failed to establish a terminal infection in an immunocompromised mouse model of systemic aspergillosis. Collectively, the pNiiA-CPR strategy enables a rapid and reliable means to directly identify, phenotypically characterize, and facilitate target-based whole cell assays to screen A. fumigatus essential genes for cognate antifungal inhibitors.

【 授权许可】

CC BY