The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases

Abstract Cryptococcosis is a fungal disease caused by C. neoformans. To adapt and survive in diverse ecological niches, including the animal host, this opportunistic pathogen relies on its ability to uptake nutrients, such as carbon, nitrogen, iron, phosphate, sulfur, and amino acids. Genetic circui...

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Autores principales:	Amanda Teixeira de Melo, Kevin Felipe Martho, Thiago Nunes Roberto, Erika S. Nishiduka, Joel Machado, Otávio J. B. Brustolini, Alexandre K. Tashima, Ana Tereza Vasconcelos, Marcelo A. Vallim, Renata C. Pascon
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/116a79f3ffc64b138ed3ac4a44ce993d
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spelling	oai:doaj.org-article:116a79f3ffc64b138ed3ac4a44ce993d2021-12-02T15:07:55ZThe regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases10.1038/s41598-019-48433-52045-2322https://doaj.org/article/116a79f3ffc64b138ed3ac4a44ce993d2019-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-48433-5https://doaj.org/toc/2045-2322Abstract Cryptococcosis is a fungal disease caused by C. neoformans. To adapt and survive in diverse ecological niches, including the animal host, this opportunistic pathogen relies on its ability to uptake nutrients, such as carbon, nitrogen, iron, phosphate, sulfur, and amino acids. Genetic circuits play a role in the response to environmental changes, modulating gene expression and adjusting the microbial metabolism to the nutrients available for the best energy usage and survival. We studied the sulfur amino acid biosynthesis and its implications on C. neoformans biology and virulence. CNAG_04798 encodes a BZip protein and was annotated as CYS3, which has been considered an essential gene. However, we demonstrated that CYS3 is not essential, in fact, its knockout led to sulfur amino acids auxotroph. Western blots and fluorescence microscopy indicated that GFP-Cys3, which is expressed from a constitutive promoter, localizes to the nucleus in rich medium (YEPD); the addition of methionine and cysteine as sole nitrogen source (SD–N + Met/Cys) led to reduced nuclear localization and protein degradation. By proteomics, we identified and confirmed physical interaction among Gpp2, Cna1, Cnb1 and GFP-Cys3. Deletion of the calcineurin and GPP2 genes in a GFP-Cys3 background demonstrated that calcineurin is required to maintain Cys3 high protein levels in YEPD and that deletion of GPP2 causes GFP-Cys3 to persist in the presence of sulfur amino acids. Global transcriptional profile of mutant and wild type by RNAseq revealed that Cys3 controls all branches of the sulfur amino acid biosynthesis, and sulfur starvation leads to induction of several amino acid biosynthetic routes. In addition, we found that Cys3 is required for virulence in Galleria mellonella animal model.Amanda Teixeira de MeloKevin Felipe MarthoThiago Nunes RobertoErika S. NishidukaJoel MachadoOtávio J. B. BrustoliniAlexandre K. TashimaAna Tereza VasconcelosMarcelo A. VallimRenata C. PasconNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-19 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Amanda Teixeira de Melo Kevin Felipe Martho Thiago Nunes Roberto Erika S. Nishiduka Joel Machado Otávio J. B. Brustolini Alexandre K. Tashima Ana Tereza Vasconcelos Marcelo A. Vallim Renata C. Pascon The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases
description	Abstract Cryptococcosis is a fungal disease caused by C. neoformans. To adapt and survive in diverse ecological niches, including the animal host, this opportunistic pathogen relies on its ability to uptake nutrients, such as carbon, nitrogen, iron, phosphate, sulfur, and amino acids. Genetic circuits play a role in the response to environmental changes, modulating gene expression and adjusting the microbial metabolism to the nutrients available for the best energy usage and survival. We studied the sulfur amino acid biosynthesis and its implications on C. neoformans biology and virulence. CNAG_04798 encodes a BZip protein and was annotated as CYS3, which has been considered an essential gene. However, we demonstrated that CYS3 is not essential, in fact, its knockout led to sulfur amino acids auxotroph. Western blots and fluorescence microscopy indicated that GFP-Cys3, which is expressed from a constitutive promoter, localizes to the nucleus in rich medium (YEPD); the addition of methionine and cysteine as sole nitrogen source (SD–N + Met/Cys) led to reduced nuclear localization and protein degradation. By proteomics, we identified and confirmed physical interaction among Gpp2, Cna1, Cnb1 and GFP-Cys3. Deletion of the calcineurin and GPP2 genes in a GFP-Cys3 background demonstrated that calcineurin is required to maintain Cys3 high protein levels in YEPD and that deletion of GPP2 causes GFP-Cys3 to persist in the presence of sulfur amino acids. Global transcriptional profile of mutant and wild type by RNAseq revealed that Cys3 controls all branches of the sulfur amino acid biosynthesis, and sulfur starvation leads to induction of several amino acid biosynthetic routes. In addition, we found that Cys3 is required for virulence in Galleria mellonella animal model.
format	article
author	Amanda Teixeira de Melo Kevin Felipe Martho Thiago Nunes Roberto Erika S. Nishiduka Joel Machado Otávio J. B. Brustolini Alexandre K. Tashima Ana Tereza Vasconcelos Marcelo A. Vallim Renata C. Pascon
author_facet	Amanda Teixeira de Melo Kevin Felipe Martho Thiago Nunes Roberto Erika S. Nishiduka Joel Machado Otávio J. B. Brustolini Alexandre K. Tashima Ana Tereza Vasconcelos Marcelo A. Vallim Renata C. Pascon
author_sort	Amanda Teixeira de Melo
title	The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases
title_short	The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases
title_full	The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases
title_fullStr	The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases
title_full_unstemmed	The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases
title_sort	regulation of the sulfur amino acid biosynthetic pathway in cryptococcus neoformans: the relationship of cys3, calcineurin, and gpp2 phosphatases
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/116a79f3ffc64b138ed3ac4a44ce993d
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The regulation of the sulfur amino acid biosynthetic pathway in Cryptococcus neoformans: the relationship of Cys3, Calcineurin, and Gpp2 phosphatases

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