Genome‐wide profiling of piggyBac transposon insertion mutants reveals loss of the F1F0 ATPase complex causes fluconazole resistance in Candida glabrata

Author:

Chow Eve W. L.1,Song Yabing23,Wang Haitao1,Xu Xiaoli1,Gao Jiaxin3,Wang Yue14ORCID

Affiliation:

1. A*STAR Infectious Diseases Labs (A*STAR ID Labs) Agency for Science and Technology Research (A*STAR) Singapore Singapore

2. School of Life Sciences Tsinghua University Beijing China

3. State Key Laboratory of Mycology, Institute of Microbiology Chinese Academy of Sciences Beijing China

4. Department of Biochemistry Yong Loo Lin School of Medicine, National University of Singapore Singapore Singapore

Abstract

AbstractInvasive candidiasis caused by non‐albicans species has been on the rise, with Candida glabrata emerging as the second most common etiological agent. Candida glabrata possesses an intrinsically lower susceptibility to azoles and an alarming propensity to rapidly develop high‐level azole resistance during treatment. In this study, we have developed an efficient piggyBac (PB) transposon‐mediated mutagenesis system in C. glabrata to conduct genome‐wide genetic screens and applied it to profile genes that contribute to azole resistance. When challenged with the antifungal drug fluconazole, PB insertion into 270 genes led to significant resistance. A large subset of these genes has a role in the mitochondria, including almost all genes encoding the subunits of the F1F0 ATPase complex. We show that deleting ATP3 or ATP22 results in increased azole resistance but does not affect susceptibility to polyenes and echinocandins. The increased azole resistance is due to increased expression of PDR1 that encodes a transcription factor known to promote drug efflux pump expression. Deleting PDR1 in the atp3Δ or atp22Δ mutant resulted in hypersensitivity to fluconazole. Our results shed light on the mechanisms contributing to azole resistance in C. glabrata. This PB transposon‐mediated mutagenesis system can significantly facilitate future genome‐wide genetic screens.

Funder

National Research Foundation Singapore

Publisher

Wiley

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