AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>

ABSTRACT Aspergillosis associated with azole-resistant Aspergillus fumigatus has a mortality rate that can approach 90% in certain patient populations. The best-understood avenue for azole resistance involves changes in the cyp51A gene that encodes the target of azole drugs, lanosterol α-14 demethyl...

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Autores principales:	Sanjoy Paul, Mark Stamnes, Grace Heredge Thomas, Hong Liu, Daisuke Hagiwara, Katsuya Gomi, Scott G. Filler, W. Scott Moye-Rowley
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	ABC transporters antifungal resistance Aspergillus fumigatus molecular genetics transcriptional regulation Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/04ab0a88ea04405a8699079ec5d9d53e
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spelling	oai:doaj.org-article:04ab0a88ea04405a8699079ec5d9d53e2021-11-15T15:55:25ZAtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>10.1128/mBio.02563-182150-7511https://doaj.org/article/04ab0a88ea04405a8699079ec5d9d53e2019-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02563-18https://doaj.org/toc/2150-7511ABSTRACT Aspergillosis associated with azole-resistant Aspergillus fumigatus has a mortality rate that can approach 90% in certain patient populations. The best-understood avenue for azole resistance involves changes in the cyp51A 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 cyp51A 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 cyp51A as well as that of an important membrane transporter protein gene called abcG1. In this work, we characterize two different mutant alleles of atrR, either an overproducing or an epitope-tagged form, causing constitutive activation of this factor. Using an epitope-tagged allele of atrR 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 abcG1 and cyp51A promoters. The ATRE in cyp51A 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. IMPORTANCE Aspergillus fumigatus is the major filamentous fungal pathogen in humans. Infections associated with A. fumigatus 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 cyp51A as an important contributor to resistance in A. fumigatus. Here, we demonstrate that the transcription factor AtrR binds to a region in the cyp51A 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 A. fumigatus.Sanjoy PaulMark StamnesGrace Heredge ThomasHong LiuDaisuke HagiwaraKatsuya GomiScott G. FillerW. Scott Moye-RowleyAmerican Society for MicrobiologyarticleABC transportersantifungal resistanceAspergillus fumigatusmolecular geneticstranscriptional regulationMicrobiologyQR1-502ENmBio, Vol 10, Iss 2 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	ABC transporters antifungal resistance Aspergillus fumigatus molecular genetics transcriptional regulation Microbiology QR1-502
spellingShingle	ABC transporters antifungal resistance Aspergillus fumigatus molecular genetics transcriptional regulation Microbiology QR1-502 Sanjoy Paul Mark Stamnes Grace Heredge Thomas Hong Liu Daisuke Hagiwara Katsuya Gomi Scott G. Filler W. Scott Moye-Rowley AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
description	ABSTRACT Aspergillosis associated with azole-resistant Aspergillus fumigatus has a mortality rate that can approach 90% in certain patient populations. The best-understood avenue for azole resistance involves changes in the cyp51A 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 cyp51A 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 cyp51A as well as that of an important membrane transporter protein gene called abcG1. In this work, we characterize two different mutant alleles of atrR, either an overproducing or an epitope-tagged form, causing constitutive activation of this factor. Using an epitope-tagged allele of atrR 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 abcG1 and cyp51A promoters. The ATRE in cyp51A 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. IMPORTANCE Aspergillus fumigatus is the major filamentous fungal pathogen in humans. Infections associated with A. fumigatus 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 cyp51A as an important contributor to resistance in A. fumigatus. Here, we demonstrate that the transcription factor AtrR binds to a region in the cyp51A 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 A. fumigatus.
format	article
author	Sanjoy Paul Mark Stamnes Grace Heredge Thomas Hong Liu Daisuke Hagiwara Katsuya Gomi Scott G. Filler W. Scott Moye-Rowley
author_facet	Sanjoy Paul Mark Stamnes Grace Heredge Thomas Hong Liu Daisuke Hagiwara Katsuya Gomi Scott G. Filler W. Scott Moye-Rowley
author_sort	Sanjoy Paul
title	AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
title_short	AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
title_full	AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
title_fullStr	AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
title_full_unstemmed	AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
title_sort	atrr is an essential determinant of azole resistance in <named-content content-type="genus-species">aspergillus fumigatus</named-content>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/04ab0a88ea04405a8699079ec5d9d53e
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AtrR Is an Essential Determinant of Azole Resistance in <named-content content-type="genus-species">Aspergillus fumigatus</named-content>

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