Functional information from clinically-derived drug resistant forms of the Candida glabrata Pdr1 transcription factor

• Published in: PLoS genetics Vol. 16; no. 8; p. e1009005
• Main Authors: Simonicova, Lucia, Moye-Rowley, W Scott
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 25.08.2020; Public Library of Science (PLoS)
• Subjects: Amino acid substitution; Amino acids; Antifungal agents; Antifungal Agents - adverse effects; Antifungal Agents - pharmacology; Azoles - adverse effects; Azoles - pharmacology; Biology and Life Sciences; Biophysics; C-Terminus; Candida; Candida glabrata; Candida glabrata - drug effects; Candida glabrata - genetics; Candida glabrata - pathogenicity; Candidiasis - drug therapy; Candidiasis - genetics; Candidiasis - microbiology; Clinical isolates; Complementarity-determining region 1; Deoxyribonucleic acid; DNA; DNA-Binding Proteins; Drug resistance; Drug Resistance, Fungal - drug effects; Drug Resistance, Fungal - genetics; Drugs; Fungal Proteins - genetics; Gene expression; Gene Expression Regulation, Fungal - drug effects; Genetic aspects; Health aspects; Humans; Microbial drug resistance; Mutation; Phenotypes; Physical Sciences; Physiology; Proteins; Research and analysis methods; Transcription activation; Transcription factors; Transcriptional Activation - drug effects; United States; United States > US; Iowa City Iowa
• ISSN: 1553-7404; 1553-7390; 1553-7404
• DOI: 10.1371/journal.pgen.1009005

Azole drugs are the most frequently used antifungal agents. The pathogenic yeast Candida glabrata acquires resistance to azole drugs via single amino acid substitution mutations eliciting a gain-of-function (GOF) hyperactive phenotype in the Pdr1 transcription factor. These GOF mutants constitutively drive high transcription of target genes such as the ATP-binding cassette transporter-encoding CDR1 locus. Previous characterization of Pdr1 has demonstrated that this factor is negatively controlled by the action of a central regulatory domain (CRD) of ~700 amino acids, in which GOF mutations are often found. Our earlier experiments demonstrated that a Pdr1 derivative in which the CRD was deleted gave rise to a transcriptional regulator that could not be maintained as the sole copy of PDR1 in the cell owing to its toxically high activity. Using a set of GOF PDR1 alleles from azole-resistant clinical isolates, we have analyzed the mechanisms acting to repress Pdr1 transcriptional activity. Our data support the view that Pdr1-dependent transactivation is mediated by a complex network of transcriptional coactivators interacting with the extreme C-terminal part of Pdr1. These coactivators include but are not limited to the Mediator component Med15A. Activity of this C-terminal domain is controlled by the CRD and requires multiple regions across the C-terminus for normal function. We also provide genetic evidence for an element within the transactivation domain that mediates the interaction of Pdr1 with coactivators on one hand while restricting Pdr1 activity on the other hand. These data indicate that GOF mutations in PDR1 block nonidentical negative inputs that would otherwise restrain Pdr1 transcriptional activation. The strong C-terminal transactivation domain of Pdr1 uses multiple different protein regions to recruit coactivators.