Klimova Natalia, Yeung Ralph, Kachurina Nadezda, Turcotte Bernard
Department of Medicine, McGill University Health Centre, McGill University, Montréal, Québec, Canada H3A 1A1.
Department of Biochemistry, McGill University Health Centre, McGill University, Montréal, Québec, Canada H3A 1A1.
G3 (Bethesda). 2014 Mar 21;4(5):931-40. doi: 10.1534/g3.113.010199.
Candida glabrata is the second most important human fungal pathogen. Despite its formal name, C. glabrata is in fact more closely related to the nonpathogenic budding yeast Saccharomyces cerevisiae. However, less is known about the biology of this pathogen. Zinc cluster proteins form a large family of transcriptional regulators involved in the regulation of numerous processes such as the control of the metabolism of sugars, amino acids, fatty acids, as well as drug resistance. The C. glabrata genome encodes 41 known or putative zinc cluster proteins, and the majority of them are uncharacterized. We have generated a panel of strains carrying individual deletions of zinc cluster genes. Using a novel approach relying on tetracycline for conditional expression in C. glabrata at the translational level, we show that only two zinc cluster genes are essential. We have performed phenotypic analysis of nonessential zinc cluster genes. Our results show that two deletion strains are thermosensitive whereas two strains are sensitive to caffeine, an inhibitor of the target of rapamycin pathway. Increased salt tolerance has been observed for eight deletion strains, whereas one strain showed reduced tolerance to salt. We have also identified a number of strains with increased susceptibility to the antifungal drugs fluconazole and ketoconazole. Interestingly, one deletion strain showed decreased susceptibility to the antifungal micafungin. In summary, we have assigned phenotypes to more than half of the zinc cluster genes in C. glabrata. Our study provides a resource that will be useful to better understand the biological role of these transcription factors.
光滑念珠菌是第二重要的人类真菌病原体。尽管其学名如此,但光滑念珠菌实际上与非致病性出芽酵母酿酒酵母关系更为密切。然而,人们对这种病原体的生物学特性了解较少。锌簇蛋白构成了一个庞大的转录调节因子家族,参与调控众多过程,如糖类、氨基酸、脂肪酸代谢的控制以及耐药性。光滑念珠菌基因组编码41种已知或推定的锌簇蛋白,其中大多数尚未被表征。我们构建了一组携带锌簇基因单个缺失的菌株。通过一种依赖四环素在光滑念珠菌中进行翻译水平条件表达的新方法,我们发现只有两个锌簇基因是必需的。我们对非必需锌簇基因进行了表型分析。我们的结果表明,两个缺失菌株对温度敏感,而两个菌株对雷帕霉素靶标途径的抑制剂咖啡因敏感。八个缺失菌株表现出耐盐性增加,而一个菌株表现出对盐的耐受性降低。我们还鉴定出一些对抗真菌药物氟康唑和酮康唑敏感性增加的菌株。有趣的是,一个缺失菌株对抗真菌药物米卡芬净的敏感性降低。总之,我们已经为光滑念珠菌中超过一半的锌簇基因赋予了表型。我们的研究提供了一种资源,将有助于更好地理解这些转录因子的生物学作用。