Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>

ABSTRACT Ergosterol (ERG) is a critical sterol in the cell membranes of fungi, and its biosynthesis is tightly regulated by 25 known enzymes along the ERG production pathway. The effects of changes in expression of each ERG biosynthesis enzyme in Saccharomyces cerevisiae were analyzed by the use of...

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Autores principales:	Somanon Bhattacharya, Brooke D. Esquivel, Theodore C. White
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Saccharomyces cerevisiae antifungal drug resistance ergosterol biosynthesis ergosterol gene overexpression ergosterol regulation stress agents Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c3f1bbde2b7e4472b74e0932c31f0805
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spelling	oai:doaj.org-article:c3f1bbde2b7e4472b74e0932c31f08052021-11-15T16:00:16ZOverexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>10.1128/mBio.01291-182150-7511https://doaj.org/article/c3f1bbde2b7e4472b74e0932c31f08052018-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01291-18https://doaj.org/toc/2150-7511ABSTRACT Ergosterol (ERG) is a critical sterol in the cell membranes of fungi, and its biosynthesis is tightly regulated by 25 known enzymes along the ERG production pathway. The effects of changes in expression of each ERG biosynthesis enzyme in Saccharomyces cerevisiae were analyzed by the use of gene deletion or plasmid-borne overexpression constructs. The strains overexpressing the ERG pathway genes were examined for changes in doubling time and responses to a variety of stress agents. In addition, ERG gene overexpression strains and ERG gene deletion strains were tested for alterations in antifungal drug susceptibility. The data show that disruptions in ergosterol biosynthesis regulation can affect a diverse set of cellular processes and can cause numerous phenotypic effects. Some of the phenotypes observed include dramatic increases in doubling times, respiratory deficiencies on glycerol media, cell wall insufficiencies on Congo red media, and disrupted ion homeostasis under iron or calcium starvation conditions. Overexpression or deletion of specific enzymes in the ERG pathway causes altered susceptibilities to a variety of classes of antifungal ergosterol inhibitors, including fluconazole, fenpropimorph, lovastatin, nystatin, amphotericin B, and terbinafine. This analysis of the effect of perturbations to the ERG pathway caused by systematic overexpression of each of the ERG pathway genes contributes significantly to the understanding of the ergosterol biosynthetic pathway and its relationship to stress response and basic biological processes. The data indicate that precise regulation of ERG genes is essential for cellular homeostasis and identify several ERG genes that could be exploited in future antifungal development efforts. IMPORTANCE A common target of antifungal drug treatment is the fungal ergosterol biosynthesis pathway. This report helps to identify ergosterol biosynthesis enzymes that have not previously been appreciated as drug targets. The effects of overexpression of each of the 25 ERG genes in S. cerevisiae were analyzed in the presence of six stress agents that target essential cellular processes (cell wall biosynthesis, protein translation, respiration, osmotic/ionic stress, and iron and calcium homeostasis), as well as six antifungal inhibitors that target ergosterol biosynthesis. The importance of identifying cell perturbations caused by gene overexpression or deletion is emphasized by the prevalence of gene expression alterations in many pathogenic and drug-resistant clinical isolates. Genes whose altered expression causes the most extensive phenotypic alterations in the presence of stressors or inhibitors have the potential to be drug targets.Somanon BhattacharyaBrooke D. EsquivelTheodore C. WhiteAmerican Society for MicrobiologyarticleSaccharomyces cerevisiaeantifungal drug resistanceergosterol biosynthesisergosterol gene overexpressionergosterol regulationstress agentsMicrobiologyQR1-502ENmBio, Vol 9, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Saccharomyces cerevisiae antifungal drug resistance ergosterol biosynthesis ergosterol gene overexpression ergosterol regulation stress agents Microbiology QR1-502
spellingShingle	Saccharomyces cerevisiae antifungal drug resistance ergosterol biosynthesis ergosterol gene overexpression ergosterol regulation stress agents Microbiology QR1-502 Somanon Bhattacharya Brooke D. Esquivel Theodore C. White Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>
description	ABSTRACT Ergosterol (ERG) is a critical sterol in the cell membranes of fungi, and its biosynthesis is tightly regulated by 25 known enzymes along the ERG production pathway. The effects of changes in expression of each ERG biosynthesis enzyme in Saccharomyces cerevisiae were analyzed by the use of gene deletion or plasmid-borne overexpression constructs. The strains overexpressing the ERG pathway genes were examined for changes in doubling time and responses to a variety of stress agents. In addition, ERG gene overexpression strains and ERG gene deletion strains were tested for alterations in antifungal drug susceptibility. The data show that disruptions in ergosterol biosynthesis regulation can affect a diverse set of cellular processes and can cause numerous phenotypic effects. Some of the phenotypes observed include dramatic increases in doubling times, respiratory deficiencies on glycerol media, cell wall insufficiencies on Congo red media, and disrupted ion homeostasis under iron or calcium starvation conditions. Overexpression or deletion of specific enzymes in the ERG pathway causes altered susceptibilities to a variety of classes of antifungal ergosterol inhibitors, including fluconazole, fenpropimorph, lovastatin, nystatin, amphotericin B, and terbinafine. This analysis of the effect of perturbations to the ERG pathway caused by systematic overexpression of each of the ERG pathway genes contributes significantly to the understanding of the ergosterol biosynthetic pathway and its relationship to stress response and basic biological processes. The data indicate that precise regulation of ERG genes is essential for cellular homeostasis and identify several ERG genes that could be exploited in future antifungal development efforts. IMPORTANCE A common target of antifungal drug treatment is the fungal ergosterol biosynthesis pathway. This report helps to identify ergosterol biosynthesis enzymes that have not previously been appreciated as drug targets. The effects of overexpression of each of the 25 ERG genes in S. cerevisiae were analyzed in the presence of six stress agents that target essential cellular processes (cell wall biosynthesis, protein translation, respiration, osmotic/ionic stress, and iron and calcium homeostasis), as well as six antifungal inhibitors that target ergosterol biosynthesis. The importance of identifying cell perturbations caused by gene overexpression or deletion is emphasized by the prevalence of gene expression alterations in many pathogenic and drug-resistant clinical isolates. Genes whose altered expression causes the most extensive phenotypic alterations in the presence of stressors or inhibitors have the potential to be drug targets.
format	article
author	Somanon Bhattacharya Brooke D. Esquivel Theodore C. White
author_facet	Somanon Bhattacharya Brooke D. Esquivel Theodore C. White
author_sort	Somanon Bhattacharya
title	Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>
title_short	Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>
title_full	Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>
title_fullStr	Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>
title_full_unstemmed	Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>
title_sort	overexpression or deletion of ergosterol biosynthesis genes alters doubling time, response to stress agents, and drug susceptibility in <italic toggle="yes">saccharomyces cerevisiae</italic>
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/c3f1bbde2b7e4472b74e0932c31f0805
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Overexpression or Deletion of Ergosterol Biosynthesis Genes Alters Doubling Time, Response to Stress Agents, and Drug Susceptibility in <italic toggle="yes">Saccharomyces cerevisiae</italic>

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