Induction of Candida albicans drug resistance genes by hybrid zinc cluster transcription factors.
The pathogenic yeast Candida albicans can develop resistance to azole antifungal drugs by overexpressing ERG11, which encodes the drug target, or the multidrug efflux pumps MDR1 and CDR1/CDR2. The constitutive upregulation of these genes is usually caused by gain-of-function mutations in the zinc cluster transcription factors Upc2, Mrr1, and Tac1, respectively. These transcription factors are also required for the induction of their target genes in drug-susceptible strains in the presence of specific stimuli. By swapping the DNA-binding domains of Mrr1, Tac1, and Upc2 we investigated if the hybrid transcription factors could activate their new target genes in response to the same signals. When Tac1 was targeted to the MDR1 and ERG11 promoters, the expression of these genes became inducible by fluphenazine. Similarly, MDR1 and CDR2 were strongly upregulated by fluconazole when Upc2 was fused to the DNA-binding domains of Mrr1 and Tac1, respectively. In contrast, Mrr1 was unable to promote gene expression in response to benomyl when it was targeted to the CDR2 and ERG11 promoters instead of the MDR1 promoter. These results suggest that Tac1 and Upc2 themselves are activated by the inducers fluphenazine and fluconazole, respectively, whereas benomyl does not activate Mrr1 itself but a coregulatory factor that is present at the promoters of Mrr1 target genes. Strains in which the expression levels of Mrr1 and Tac1 target genes were controlled by Upc2 exhibited increased fluconazole resistance, demonstrating that the ability to efficiently upregulate the expression of efflux pumps in the presence of the drug results in enhanced intrinsic fluconazole resistance.
Institut für Molekulare Infektionsbiologie, Universität Würzburg, Würzburg, Germany.
Sabrina
S
Schneider
Institut für Molekulare Infektionsbiologie, Universität Würzburg, Würzburg, Germany joachim.morschhaeuser@uni-wuerzburg.de.
Joachim
J
Morschhäuser
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics; D000935:Antifungal Agents / Q000494:pharmacology*; D001665:Binding Sites; D002176:Candida albicans / Q000187:drug effects* / Q000235:genetics*; D025141:Drug Resistance, Fungal / Q000187:drug effects / Q000235:genetics*; D015725:Fluconazole / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000935:Antifungal Agents; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D015725:Fluconazole; D015032:Zinc