title: Candida glabrata drug:H+ antiporter CgQdr2 confers imidazole drug resistance, being activated by transcription factor CgPdr1.
abstract: The widespread emergence of antifungal drug resistance poses a severe clinical problem. Though predicted to play a role in this phenomenon, the drug:H(+) antiporters (DHA) of the major facilitator superfamily have largely escaped characterization in pathogenic yeasts. This work describes the first DHA from the pathogenic yeast Candida glabrata reported to be involved in antifungal drug resistance, the C. glabrata QDR2 (CgQDR2) gene (ORF CAGL0G08624g). The expression of CgQDR2 in C. glabrata was found to confer resistance to the antifungal drugs miconazole, tioconazole, clotrimazole, and ketoconazole. By use of a green fluorescent protein (GFP) fusion, the CgQdr2 protein was found to be targeted to the plasma membrane in C. glabrata. In agreement with these observations, CgQDR2 expression was found to decrease the intracellular accumulation of radiolabeled clotrimazole in C. glabrata and to play a role in the extrusion of this antifungal from preloaded cells. Interestingly, the functional heterologous expression of CgQDR2 in the model yeast Saccharomyces cerevisiae further confirmed the role of this gene as a multidrug resistance determinant: its expression was able to complement the susceptibility phenotype exhibited by its S. cerevisiae homologue, QDR2, in the presence of imidazoles and of the antimalarial and antiarrhythmic drug quinidine. In contrast to the findings reported for Qdr2, CgQdr2 expression does not contribute to the ability of yeast to grow under K(+)-limiting conditions. Interestingly, CgQDR2 transcript levels were seen to be upregulated in C. glabrata cells challenged with clotrimazole or quinidine. This upregulation was found to depend directly on the transcription factor CgPdr1, the major regulator of multidrug resistance in this pathogenic yeast, which has also been found to be a determinant of quinidine and clotrimazole resistance in C. glabrata.
authors: Department of Bioengineering, Instituto Superior Técnico, Technical University of Lisbon, Lisbon, Portugal.
authors: Catarina
authors: C
authors: Costa
authors: Carla
authors: C
authors: Pires
authors: Tânia R
authors: TR
authors: Cabrito
authors: Adeline
authors: A
authors: Renaudin
authors: Michiyo
authors: M
authors: Ohno
authors: Hiroji
authors: H
authors: Chibana
authors: Isabel
authors: I
authors: Sá-Correia
authors: Miguel C
authors: MC
authors: Teixeira
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000935:Antifungal Agents / Q000494:pharmacology*; D017920:Antiporters / Q000378:metabolism*; D041221:Candida glabrata / Q000187:drug effects* / Q000378:metabolism; D003022:Clotrimazole / Q000494:pharmacology; D025141:Drug Resistance, Fungal / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D049452:Green Fluorescent Proteins; D007093:Imidazoles / Q000494:pharmacology*; D007654:Ketoconazole / Q000494:pharmacology; D008825:Miconazole / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents; D017920:Antiporters; D007093:Imidazoles; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D008825:Miconazole; D003022:Clotrimazole; D007654:Ketoconazole; C020527:tioconazole
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