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Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay

New antifungal drugs are urgently needed due to the currently limited selection, the emergence of drug resistance, and the toxicity of several commonly used drugs. To identify drug leads, we screened small molecules using a Saccharomyces cerevisiae reporter bioassay in which S. cerevisiae heterologo...

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Autores principales: Tebbets, Brad, Stewart, Douglas, Lawry, Stephanie, Nett, Jeniel, Nantel, Andre, Andes, David, Klein, Bruce S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3344848/
https://www.ncbi.nlm.nih.gov/pubmed/22574132
http://dx.doi.org/10.1371/journal.pone.0036021
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author Tebbets, Brad
Stewart, Douglas
Lawry, Stephanie
Nett, Jeniel
Nantel, Andre
Andes, David
Klein, Bruce S.
author_facet Tebbets, Brad
Stewart, Douglas
Lawry, Stephanie
Nett, Jeniel
Nantel, Andre
Andes, David
Klein, Bruce S.
author_sort Tebbets, Brad
collection PubMed
description New antifungal drugs are urgently needed due to the currently limited selection, the emergence of drug resistance, and the toxicity of several commonly used drugs. To identify drug leads, we screened small molecules using a Saccharomyces cerevisiae reporter bioassay in which S. cerevisiae heterologously expresses Hik1, a group III hybrid histidine kinase (HHK) from Magnaporthe grisea. Group III HHKs are integral in fungal cell physiology, and highly conserved throughout this kingdom; they are absent in mammals, making them an attractive drug target. Our screen identified compounds 13 and 33, which showed robust activity against numerous fungal genera including Candida spp., Cryptococcus spp. and molds such as Aspergillus fumigatus and Rhizopus oryzae. Drug-resistant Candida albicans from patients were also highly susceptible to compounds 13 and 33. While the compounds do not act directly on HHKs, microarray analysis showed that compound 13 induced transcripts associated with oxidative stress, and compound 33, transcripts linked with heavy metal stress. Both compounds were highly active against C. albicans biofilm, in vitro and in vivo, and exerted synergy with fluconazole, which was inactive alone. Thus, we identified potent, broad-spectrum antifungal drug leads from a small molecule screen using a high-throughput, S. cerevisiae reporter bioassay.
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spelling pubmed-33448482012-05-09 Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay Tebbets, Brad Stewart, Douglas Lawry, Stephanie Nett, Jeniel Nantel, Andre Andes, David Klein, Bruce S. PLoS One Research Article New antifungal drugs are urgently needed due to the currently limited selection, the emergence of drug resistance, and the toxicity of several commonly used drugs. To identify drug leads, we screened small molecules using a Saccharomyces cerevisiae reporter bioassay in which S. cerevisiae heterologously expresses Hik1, a group III hybrid histidine kinase (HHK) from Magnaporthe grisea. Group III HHKs are integral in fungal cell physiology, and highly conserved throughout this kingdom; they are absent in mammals, making them an attractive drug target. Our screen identified compounds 13 and 33, which showed robust activity against numerous fungal genera including Candida spp., Cryptococcus spp. and molds such as Aspergillus fumigatus and Rhizopus oryzae. Drug-resistant Candida albicans from patients were also highly susceptible to compounds 13 and 33. While the compounds do not act directly on HHKs, microarray analysis showed that compound 13 induced transcripts associated with oxidative stress, and compound 33, transcripts linked with heavy metal stress. Both compounds were highly active against C. albicans biofilm, in vitro and in vivo, and exerted synergy with fluconazole, which was inactive alone. Thus, we identified potent, broad-spectrum antifungal drug leads from a small molecule screen using a high-throughput, S. cerevisiae reporter bioassay. Public Library of Science 2012-05-04 /pmc/articles/PMC3344848/ /pubmed/22574132 http://dx.doi.org/10.1371/journal.pone.0036021 Text en Tebbets et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Tebbets, Brad
Stewart, Douglas
Lawry, Stephanie
Nett, Jeniel
Nantel, Andre
Andes, David
Klein, Bruce S.
Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay
title Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay
title_full Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay
title_fullStr Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay
title_full_unstemmed Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay
title_short Identification and Characterization of Antifungal Compounds Using a Saccharomyces cerevisiae Reporter Bioassay
title_sort identification and characterization of antifungal compounds using a saccharomyces cerevisiae reporter bioassay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3344848/
https://www.ncbi.nlm.nih.gov/pubmed/22574132
http://dx.doi.org/10.1371/journal.pone.0036021
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