Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses

ABSTRACT Alternative splicing (AS)—a process by which a single gene gives rise to different protein isoforms in eukaryotes—has been implicated in many basic cellular processes, but little is known about its role in drug resistance and fungal pathogenesis. The most common human fungal pathogen, Candi...

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Autores principales:	Suraya Muzafar, Ravi Datta Sharma, Abdul Haseeb Shah, Naseem A. Gaur, Ujjaini Dasgupta, Neeraj Chauhan, Rajendra Prasad
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Candida albicans alternative splicing SOD3 amphotericin B menadione Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/4d18dc212f064722998e93d1459c336d
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spelling	oai:doaj.org-article:4d18dc212f064722998e93d1459c336d2021-11-15T15:30:51ZIdentification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses10.1128/mSphere.00608-202379-5042https://doaj.org/article/4d18dc212f064722998e93d1459c336d2020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00608-20https://doaj.org/toc/2379-5042ABSTRACT Alternative splicing (AS)—a process by which a single gene gives rise to different protein isoforms in eukaryotes—has been implicated in many basic cellular processes, but little is known about its role in drug resistance and fungal pathogenesis. The most common human fungal pathogen, Candida albicans, has introns in 4 to 6% of its genes, the functions of which remain largely unknown. Here, we report AS regulating drug resistance in C. albicans. Comparative RNA-sequencing of two different sets of sequential, isogenic azole-sensitive and -resistant isolates of C. albicans revealed differential expression of splice isoforms of 14 genes. One of these was the superoxide dismutase gene SOD3, which contains a single intron. The sod3Δ/Δ mutant was susceptible to the antifungals amphotericin B (AMB) and menadione (MND). While AMB susceptibility was rescued by overexpression of both the spliced and unspliced SOD3 isoforms, only the spliced isoform could overcome MND susceptibility, demonstrating the functional relevance of this splicing in developing drug resistance. Furthermore, unlike AMB, MND inhibits SOD3 splicing and acts as a splicing inhibitor. Consistent with these observations, MND exposure resulted in increased levels of unspliced SOD3 isoform that are unable to scavenge reactive oxygen species (ROS), resulting in increased drug susceptibility. Collectively, these observations suggest that AS is a novel mechanism for stress adaptation and overcoming drug susceptibility in C. albicans. IMPORTANCE The emergence of resistance in Candida albicans, an opportunistic pathogen, against the commonly used antifungals is becoming a major obstacle in its treatment. The necessity to identify new drug targets demands fundamental insights into the mechanisms used by this organism to develop drug resistance. C. albicans has introns in 4 to 6% of its genes, the functions of which remain largely unknown. Using the RNA-sequencing data from isogenic pairs of azole-sensitive and -resistant isolates of C. albicans, here, we show how C. albicans uses modulations in mRNA splicing to overcome antifungal drug stress.Suraya MuzafarRavi Datta SharmaAbdul Haseeb ShahNaseem A. GaurUjjaini DasguptaNeeraj ChauhanRajendra PrasadAmerican Society for MicrobiologyarticleCandida albicansalternative splicingSOD3amphotericin BmenadioneMicrobiologyQR1-502ENmSphere, Vol 5, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Candida albicans alternative splicing SOD3 amphotericin B menadione Microbiology QR1-502
spellingShingle	Candida albicans alternative splicing SOD3 amphotericin B menadione Microbiology QR1-502 Suraya Muzafar Ravi Datta Sharma Abdul Haseeb Shah Naseem A. Gaur Ujjaini Dasgupta Neeraj Chauhan Rajendra Prasad Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses
description	ABSTRACT Alternative splicing (AS)—a process by which a single gene gives rise to different protein isoforms in eukaryotes—has been implicated in many basic cellular processes, but little is known about its role in drug resistance and fungal pathogenesis. The most common human fungal pathogen, Candida albicans, has introns in 4 to 6% of its genes, the functions of which remain largely unknown. Here, we report AS regulating drug resistance in C. albicans. Comparative RNA-sequencing of two different sets of sequential, isogenic azole-sensitive and -resistant isolates of C. albicans revealed differential expression of splice isoforms of 14 genes. One of these was the superoxide dismutase gene SOD3, which contains a single intron. The sod3Δ/Δ mutant was susceptible to the antifungals amphotericin B (AMB) and menadione (MND). While AMB susceptibility was rescued by overexpression of both the spliced and unspliced SOD3 isoforms, only the spliced isoform could overcome MND susceptibility, demonstrating the functional relevance of this splicing in developing drug resistance. Furthermore, unlike AMB, MND inhibits SOD3 splicing and acts as a splicing inhibitor. Consistent with these observations, MND exposure resulted in increased levels of unspliced SOD3 isoform that are unable to scavenge reactive oxygen species (ROS), resulting in increased drug susceptibility. Collectively, these observations suggest that AS is a novel mechanism for stress adaptation and overcoming drug susceptibility in C. albicans. IMPORTANCE The emergence of resistance in Candida albicans, an opportunistic pathogen, against the commonly used antifungals is becoming a major obstacle in its treatment. The necessity to identify new drug targets demands fundamental insights into the mechanisms used by this organism to develop drug resistance. C. albicans has introns in 4 to 6% of its genes, the functions of which remain largely unknown. Using the RNA-sequencing data from isogenic pairs of azole-sensitive and -resistant isolates of C. albicans, here, we show how C. albicans uses modulations in mRNA splicing to overcome antifungal drug stress.
format	article
author	Suraya Muzafar Ravi Datta Sharma Abdul Haseeb Shah Naseem A. Gaur Ujjaini Dasgupta Neeraj Chauhan Rajendra Prasad
author_facet	Suraya Muzafar Ravi Datta Sharma Abdul Haseeb Shah Naseem A. Gaur Ujjaini Dasgupta Neeraj Chauhan Rajendra Prasad
author_sort	Suraya Muzafar
title	Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses
title_short	Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses
title_full	Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses
title_fullStr	Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses
title_full_unstemmed	Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses
title_sort	identification of genomewide alternative splicing events in sequential, isogenic clinical isolates of <named-content content-type="genus-species">candida albicans</named-content> reveals a novel mechanism of drug resistance and tolerance to cellular stresses
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/4d18dc212f064722998e93d1459c336d
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Identification of Genomewide Alternative Splicing Events in Sequential, Isogenic Clinical Isolates of <named-content content-type="genus-species">Candida albicans</named-content> Reveals a Novel Mechanism of Drug Resistance and Tolerance to Cellular Stresses

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