title: Phenotypic analysis of a family of transcriptional regulators, the zinc cluster proteins, in the human fungal pathogen Candida glabrata.
abstract: 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.
authors.affiliation: Department of Medicine, McGill University Health Centre, McGill University, Montréal, Québec, Canada H3A 1A1.
authors.forename: Natalia
authors.initials: N
authors.lastname: Klimova
authors.affiliation: Department of Biochemistry, McGill University Health Centre, McGill University, Montréal, Québec, Canada H3A 1A1.
authors.forename: Ralph
authors.initials: R
authors.lastname: Yeung
authors.affiliation: Department of Medicine, McGill University Health Centre, McGill University, Montréal, Québec, Canada H3A 1A1.
authors.forename: Nadezda
authors.initials: N
authors.lastname: Kachurina
authors.affiliation: 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 Department of Microbiology and Immunology, McGill University Health Centre, McGill University, Montréal, Québec, Canada H3A 1A1 bernard.turcotte@mcgill.ca.
authors.forename: Bernard
authors.initials: B
authors.lastname: Turcotte
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000494:pharmacology; D002110:Caffeine / Q000494:pharmacology; D041221:Candida glabrata / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D017353:Gene Deletion; D023061:Gene Order; D006801:Humans; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D010641:Phenotype; D054730:Protein Interaction Domains and Motifs; D055049:Salt Tolerance / Q000235:genetics; D016415:Sequence Alignment; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000935:Antifungal Agents; D014157:Transcription Factors; D002110:Caffeine
keywords: Candida glabrata; drug resistance; phenotypic analysis; transcriptional regulators; zinc cluster proteins