Uncontrolled transposition following RNAi loss causes hypermutation and antifungal drug resistance in clinical isolates of Cryptococcus neoformans.

Autor: Priest SJ; Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA., Yadav V; Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA., Roth C; Department of Biology, Duke University, Durham, NC, USA.; University Program in Genetics and Genomics, Duke University, Durham, NC, USA., Dahlmann TA; Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, Bochum, Germany., Kück U; Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, Bochum, Germany., Magwene PM; Department of Biology, Duke University, Durham, NC, USA., Heitman J; Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC, USA. heitm001@duke.edu.
Jazyk: angličtina
Zdroj: Nature microbiology [Nat Microbiol] 2022 Aug; Vol. 7 (8), pp. 1239-1251. Date of Electronic Publication: 2022 Aug 02.
DOI: 10.1038/s41564-022-01183-z
Abstrakt: Cryptococcus neoformans infections cause approximately 15% of AIDS-related deaths owing to a combination of limited antifungal therapies and drug resistance. A collection of clinical and environmental C. neoformans isolates were assayed for increased mutation rates via fluctuation analysis, and we identified two hypermutator C. neoformans clinical isolates with increased mutation rates when exposed to the combination of rapamycin and FK506. Sequencing of drug target genes found that Cnl1 transposon insertions conferred the majority of resistance to rapamycin and FK506 and could also independently cause resistance to 5-fluoroorotic acid and the clinically relevant antifungal 5-flucytosine. Whole-genome sequencing revealed both hypermutator genomes harbour a nonsense mutation in the RNA-interference component ZNF3 and hundreds of Cnl1 elements organized into massive subtelomeric arrays on each of the fourteen chromosomes. Quantitative trait locus mapping in 28 progeny derived from a cross between a hypermutator and wild-type identified a locus associated with hypermutation that included znf3. CRISPR editing of the znf3 nonsense mutation abolished hypermutation and restored small-interfering-RNA production. We conclude that hypermutation and drug resistance in these clinical isolates result from RNA-interference loss and accumulation of Cnl1 elements.
(© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)
Databáze: MEDLINE
Externí odkaz: