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AtrR Is an Essential Determinant of Azole Resistance in Aspergillus fumigatus
Journal article   Open access   Peer reviewed

AtrR Is an Essential Determinant of Azole Resistance in Aspergillus fumigatus

Sanjoy Paul, Mark Stamnes, Grace Heredge Thomas, Hong Liu, Daisuke Hagiwara, Katsuya Gomi, Scott G Filler and W Scott Moye-Rowley
mBio, Vol.10(2), p.e02563-18
03/12/2019
DOI: 10.1128/mBio.02563-18
PMCID: PMC6414702
PMID: 30862750
url
https://doi.org/10.1128/mBio.02563-18View
Published (Version of record) Open Access

Abstract

Aspergillosis associated with azole-resistant has a mortality rate that can approach 90% in certain patient populations. The best-understood avenue for azole resistance involves changes in the gene that encodes the target of azole drugs, lanosterol α-14 demethylase. The most common azole resistance allele currently described is a linked change corresponding to a change in the coding sequence of and a duplication of a 34-bp region in the promoter leading to a tandem repeat (TR). Our previous studies identified a positively acting transcription factor called AtrR that binds to the promoter of as well as that of an important membrane transporter protein gene called In this work, we characterize two different mutant alleles of , either an overproducing or an epitope-tagged form, causing constitutive activation of this factor. Using an epitope-tagged allele of for chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-seq), the genomic binding sites for AtrR were determined. Close to 900 genes were found to have an AtrR response element (ATRE) in their promoter regions. Transcriptome evaluation by RNA sequencing (RNA-seq) indicated that both alleles led to elevated transcription of a subset of target genes. An electrophoretic mobility shift assay and DNase I protection mapping localized the ATREs in both the and promoters. The ATRE in was located within the 34-bp repeat element. Virulence in a murine model was compromised when AtrR was either deleted or overproduced, indicating that the proper dosage of this factor is key for pathogenesis. is the major filamentous fungal pathogen in humans. Infections associated with are often treated with azole drugs, but resistance to these antifungal agents is increasing. Mortality from aspergillosis associated with azole-resistant fungi is extremely high. Previous work has identified transcriptional control of the azole drug target-encoding gene as an important contributor to resistance in Here, we demonstrate that the transcription factor AtrR binds to a region in the promoter that is associated with alleles of this gene conferring clinically important azole resistance. Using high-throughput genomic technologies, we also uncover a large suite of target genes controlled by AtrR. These data indicate that AtrR coordinately regulates many different processes involved in drug resistance, metabolism, and virulence. Our new understanding of AtrR function provides important new insight into the pathogenesis of .
Antifungal Agents - pharmacology Aspergillus fumigatus - drug effects Aspergillus fumigatus - genetics Azoles - pharmacology Chromatin Immunoprecipitation DNA, Fungal - metabolism Drug Resistance, Fungal Electrophoretic Mobility Shift Assay Fungal Proteins - genetics Fungal Proteins - metabolism Gene Deletion Gene Expression Profiling Gene Expression Regulation, Fungal Mutation Promoter Regions, Genetic Protein Binding Regulon Sequence Analysis, DNA Transcription Factors - genetics Transcription Factors - metabolism

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