Schwarzmüller Tobias, Ma Biao, Hiller Ekkehard, Istel Fabian, Tscherner Michael, Brunke Sascha, Ames Lauren, Firon Arnaud, Green Brian, Cabral Vitor, Marcet-Houben Marina, Jacobsen Ilse D, Quintin Jessica, Seider Katja, Frohner Ingrid, Glaser Walter, Jungwirth Helmut, Bachellier-Bassi Sophie, Chauvel Murielle, Zeidler Ute, Ferrandon Dominique, Gabaldón Toni, Hube Bernhard, d'Enfert Christophe, Rupp Steffen, Cormack Brendan, Haynes Ken, Kuchler Karl
Medical University Vienna, Max F. Perutz Laboratories, Department of Medical Biochemistry, Vienna, Austria.
Department of Microbiology, Imperial College London, London, United Kingdom; Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
PLoS Pathog. 2014 Jun 19;10(6):e1004211. doi: 10.1371/journal.ppat.1004211. eCollection 2014 Jun.
The opportunistic fungal pathogen Candida glabrata is a frequent cause of candidiasis, causing infections ranging from superficial to life-threatening disseminated disease. The inherent tolerance of C. glabrata to azole drugs makes this pathogen a serious clinical threat. To identify novel genes implicated in antifungal drug tolerance, we have constructed a large-scale C. glabrata deletion library consisting of 619 unique, individually bar-coded mutant strains, each lacking one specific gene, all together representing almost 12% of the genome. Functional analysis of this library in a series of phenotypic and fitness assays identified numerous genes required for growth of C. glabrata under normal or specific stress conditions, as well as a number of novel genes involved in tolerance to clinically important antifungal drugs such as azoles and echinocandins. We identified 38 deletion strains displaying strongly increased susceptibility to caspofungin, 28 of which encoding proteins that have not previously been linked to echinocandin tolerance. Our results demonstrate the potential of the C. glabrata mutant collection as a valuable resource in functional genomics studies of this important fungal pathogen of humans, and to facilitate the identification of putative novel antifungal drug target and virulence genes.
机会性真菌病原体光滑念珠菌是念珠菌病的常见病因,可引发从浅表感染到危及生命的播散性疾病等各种感染。光滑念珠菌对唑类药物的固有耐受性使其成为严重的临床威胁。为了鉴定与抗真菌药物耐受性相关的新基因,我们构建了一个大规模的光滑念珠菌缺失文库,该文库由619个独特的、带有个体条形码的突变菌株组成,每个菌株缺失一个特定基因,总共代表了近12%的基因组。在一系列表型和适应性分析中对该文库进行功能分析,确定了光滑念珠菌在正常或特定应激条件下生长所需的众多基因,以及一些与对临床上重要的抗真菌药物(如唑类和棘白菌素)的耐受性相关的新基因。我们鉴定出38个对卡泊芬净敏感性显著增加的缺失菌株,其中28个编码的蛋白质以前未与棘白菌素耐受性相关联。我们的结果证明了光滑念珠菌突变体库作为这种重要人类真菌病原体功能基因组学研究的宝贵资源的潜力,并有助于鉴定假定的新型抗真菌药物靶点和毒力基因。