Regulation of the CgPdr1 transcription factor from the pathogen Candida glabrata.
Candida glabrata is an opportunistic human pathogen that is increasingly associated with candidemia, owing in part to the intrinsic and acquired high tolerance the organism exhibits for the important clinical antifungal drug fluconazole. This elevated fluconazole resistance often develops through gain-of-function mutations in the zinc cluster-containing transcriptional regulator C. glabrata Pdr1 (CgPdr1). CgPdr1 induces the expression of an ATP-binding cassette (ABC) transporter-encoding gene, CgCDR1. Saccharomyces cerevisiae has two CgPdr1 homologues called ScPdr1 and ScPdr3. These factors control the expression of an ABC transporter-encoding gene called ScPDR5, which encodes a homologue of CgCDR1. Loss of the mitochondrial genome (ρ(0) cell) or overexpression of the mitochondrial enzyme ScPsd1 induces ScPDR5 expression in a strictly ScPdr3-dependent fashion. ScPdr3 requires the presence of a transcriptional Mediator subunit called Gal11 (Med15) to fully induce ScPDR5 transcription in response to ρ(0) signaling. ScPdr1 does not respond to either ρ(0) signals or ScPsd1 overproduction. In this study, we employed transcriptional fusions between CgPdr1 target promoters, like CgCDR1, to demonstrate that CgPdr1 stimulates gene expression via binding to elements called pleiotropic drug response elements (PDREs). Deletion mapping and electrophoretic mobility shift assays demonstrated that a single PDRE in the CgCDR1 promoter was capable of supporting ρ(0)-induced gene expression. Removal of one of the two ScGal11 homologues from C. glabrata caused a major defect in drug-induced expression of CgCDR1 but had a quantitatively minor effect on ρ(0)-stimulated transcription. These data demonstrate that CgPdr1 appears to combine features of ScPdr1 and ScPdr3 to produce a transcription factor with chimeric regulatory properties.
Department of Molecular Physiology and Biophysics, Carver College of Medicine, 6-530 Bowen Science Bldg., University of Iowa, Iowa City, IA 52242, USA.
Sanjoy
S
Paul
Jennifer A
JA
Schmidt
W Scott
WS
Moye-Rowley
D001665:Binding Sites; D041221:Candida glabrata / Q000502:physiology*; D002262:Carboxy-Lyases / Q000096:biosynthesis; D049109:Cell Proliferation / Q000187:drug effects; D025141:Drug Resistance, Fungal / Q000235:genetics; D015725:Fluconazole / Q000494:pharmacology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D006801:Humans; D056892:Mediator Complex / Q000235:genetics; D008928:Mitochondria / Q000201:enzymology / Q000235:genetics; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D005656:Fungal Proteins; D056892:Mediator Complex; D014157:Transcription Factors; D015725:Fluconazole; D002262:Carboxy-Lyases; C022353:phosphatidylserine decarboxylase