TY - JOUR

T1 - Induction of Aspergillus fumigatus zinc cluster transcription factor OdrA/Mdu2 provides combined cellular responses for oxidative stress protection and multiple antifungal drug resistance

AU - Sasse, Christoph

AU - Bastakis, Emmanouil

AU - Bakti, Fruzsina

AU - Höfer, Annalena M.

AU - Zangl, Isabella

AU - Schüller, Christoph

AU - Köhler, Anna M.

AU - Gerke, Jennifer

AU - Krappmann, Sven

AU - Finkernagel, Florian

AU - Harting, Rebekka

AU - Strauss, Joseph

AU - Heimel, Kai

AU - Braus, Gerhard H.

N1 - Funding Information: This research has been funded by the Deutsche Forschungsgemeinschaft (DFG) (DFG1502/19-1 and SFB860 to G.H.B.), by the University of Göttingen für wissenschaftlichen Nachwuchs,” Innenauftrag, Nr.: 3917533 to C.S.), and the GFF Lower Austria Science Fund “Bioactive Microbial Metabolites” (K3-G-2/08-2020 to J.S.). We acknowledge support by the Open Access Publication Funds of University of Göttingen.

PY - 2023/11/20

Y1 - 2023/11/20

N2 - Zinc cluster transcription factors (Zcfs) encoding zcf genes are exclusive for fungi and required for multiple cellular processes including metabolism and development. Genome-wide screening of 228 zinc cluster transcription factor encoding genes by overexpression in Aspergillus fumigatus revealed 11 genes which provided increased tolerance to the broadly applied azole voriconazole or to the polyene amphotericin B or to both. These include four oxidative stress and drug resistance genes (odrA-D) genes encoding factors, which provide broad cellular stress protection. Thereby, the corresponding fungal OdrA/Mdu2- and AtrR/OdrD-dependent genetic networks are interconnected. OdrA/Mdu2 activates atrR/odrD transcription by direct binding to the promoter, whereas AtrR/OdrD functions as repressor of odrA/mdu2 expression. odrA/ mdu2 overexpression provides combined resistance to amphotericin B, voriconazole, itraconazole, and reactive oxygen species generated by menadione. OdrA/Mdu2-mediated itraconazole resistance is evoked by direct regulation of the transporter encoding gene mdr1. Oxidative stress-inducing substances like amphotericin B and menadione promote OdrA/Mdu2 accumulation in the nucleus to regulate stress response genes like mdr1 and the putative glutathione-S-transferase encoding gene gstD. The expression levels and external stress conditions fostering nuclear accumulation of OdrA/Mdu2 determine the regulation of the target genes. Hence, OdrA/Mdu2 provides a combined adaptation strategy for survival in nature or within a potential host, where this fungus represents the most common agent for human mold pneumonia worldwide. The OdrA/ Mdu2 controlled genetic network highlights the tight connection between oxidative stress response and antifungal drug adaption to secure A. fumigatus survival in various hostile environments. IMPORTANCE An overexpression screen of 228 zinc cluster transcription factor encoding genes of A. fumigatus revealed 11 genes conferring increased tolerance to antifungal drugs. Out of these, four oxidative stress and drug tolerance transcription factor encoding odr genes increased tolerance to oxidative stress and antifungal drugs when overexpressed. This supports a correlation between oxidative stress response and antifungal drug tolerance in A. fumigatus. OdrA/Mdu2 is required for the cross-tolerance between azoles, polyenes, and oxidative stress and activates genes for detoxification. Under oxidative stress conditions or when overexpressed, OdrA/Mdu2 accumulates in the nucleus and activates detoxifying genes by direct binding at their promoters, as we describe with the mdr1 gene encoding an itraconazole specific efflux pump. Finally, this work gives new insights about drug and stress resistance in the opportunistic pathogenic fungus A. fumigatus.

AB - Zinc cluster transcription factors (Zcfs) encoding zcf genes are exclusive for fungi and required for multiple cellular processes including metabolism and development. Genome-wide screening of 228 zinc cluster transcription factor encoding genes by overexpression in Aspergillus fumigatus revealed 11 genes which provided increased tolerance to the broadly applied azole voriconazole or to the polyene amphotericin B or to both. These include four oxidative stress and drug resistance genes (odrA-D) genes encoding factors, which provide broad cellular stress protection. Thereby, the corresponding fungal OdrA/Mdu2- and AtrR/OdrD-dependent genetic networks are interconnected. OdrA/Mdu2 activates atrR/odrD transcription by direct binding to the promoter, whereas AtrR/OdrD functions as repressor of odrA/mdu2 expression. odrA/ mdu2 overexpression provides combined resistance to amphotericin B, voriconazole, itraconazole, and reactive oxygen species generated by menadione. OdrA/Mdu2-mediated itraconazole resistance is evoked by direct regulation of the transporter encoding gene mdr1. Oxidative stress-inducing substances like amphotericin B and menadione promote OdrA/Mdu2 accumulation in the nucleus to regulate stress response genes like mdr1 and the putative glutathione-S-transferase encoding gene gstD. The expression levels and external stress conditions fostering nuclear accumulation of OdrA/Mdu2 determine the regulation of the target genes. Hence, OdrA/Mdu2 provides a combined adaptation strategy for survival in nature or within a potential host, where this fungus represents the most common agent for human mold pneumonia worldwide. The OdrA/ Mdu2 controlled genetic network highlights the tight connection between oxidative stress response and antifungal drug adaption to secure A. fumigatus survival in various hostile environments. IMPORTANCE An overexpression screen of 228 zinc cluster transcription factor encoding genes of A. fumigatus revealed 11 genes conferring increased tolerance to antifungal drugs. Out of these, four oxidative stress and drug tolerance transcription factor encoding odr genes increased tolerance to oxidative stress and antifungal drugs when overexpressed. This supports a correlation between oxidative stress response and antifungal drug tolerance in A. fumigatus. OdrA/Mdu2 is required for the cross-tolerance between azoles, polyenes, and oxidative stress and activates genes for detoxification. Under oxidative stress conditions or when overexpressed, OdrA/Mdu2 accumulates in the nucleus and activates detoxifying genes by direct binding at their promoters, as we describe with the mdr1 gene encoding an itraconazole specific efflux pump. Finally, this work gives new insights about drug and stress resistance in the opportunistic pathogenic fungus A. fumigatus.

KW - Aspergillus fumigatus

KW - atrR

KW - drug tolerance

KW - mdr1

KW - OdrA/Mdu2

KW - oxidative stress

KW - zinc cluster transcription factors

UR - http://www.scopus.com/inward/record.url?scp=85183693047&partnerID=8YFLogxK

U2 - 10.1128/mbio.02628-23

DO - 10.1128/mbio.02628-23

M3 - Article

C2 - 37982619

AN - SCOPUS:85183693047

VL - 14

JO - MBIO

JF - MBIO

SN - 2161-2129

IS - 6

M1 - 0262823

ER -