The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/38116 http://dx.doi.org/10.1016/j.fgb.2014.06.003 |
Resumo: | Fungal infections are often difficult to treat due to the inherent similarities between fungal and animal cells and the resulting host toxicity from many antifungal compounds. Cryptococcus neoformans is an opportunistic fungal pathogen of humans that causes life-threatening disease, primarily in immunocompromised patients. Since antifungal therapy for this microorganism is limited, many investigators have explored novel drug targets aim at virulence factors, such as the ability to grow at mammalian physiological temperature (37 degrees C). To address this issue, we used the Agrobacterium tumefaciens gene delivery system to create a random insertion mutagenesis library that was screened for altered growth at elevated temperatures. Among several mutants unable to grow at 37 degrees C, we explored one bearing an interruption in the URA4 gene. This gene encodes dihydroorotase (DHOase) that is involved in the de nova synthesis of pyrimidine ribonucleotides. Loss of the C. neoformans Ura4 protein, by targeted gene interruption, resulted in an expected uracil/uridine auxotrophy and an unexpected high temperature growth defect. in addition, the ura4 mutant displayed phenotypic defects in other prominent virulence factors (melanin, capsule and phospholipase) and reduced stress response compared to wild type and reconstituted strains. Accordingly, this mutant had a decreased survival rate in macrophages and attenuated virulence in a murine model of cryptococcal infection. Quantitative PCR analysis suggests that this biosynthetic pathway is induced during the transition from 30 degrees C to 37 degrees C, and that transcriptional regulation of de nova and salvage pyrimidine pathway are under the control of the Ura4 protein. (C) 2014 the Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-SA license (http://creativecommons.org/licenses/by-nc-sa/3.0/). |
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Gontijo, Fabiano Assis de [UNIFESP]Pascon, Renata Castiglioni [UNIFESP]Fernandes, LarissaMachado Junior, Joel [UNIFESP]Alspaugh, J. AndrewVallim, Marcelo Afonso [UNIFESP]Universidade Federal de São Paulo (UNIFESP)Universidade de Brasília (UnB)Duke Univ2016-01-24T14:37:45Z2016-01-24T14:37:45Z2014-09-01Fungal Genetics and Biology. San Diego: Academic Press Inc Elsevier Science, v. 70, p. 12-23, 2014.1087-1845http://repositorio.unifesp.br/handle/11600/38116http://dx.doi.org/10.1016/j.fgb.2014.06.003WOS000341617000003.pdf10.1016/j.fgb.2014.06.003WOS:000341617000003Fungal infections are often difficult to treat due to the inherent similarities between fungal and animal cells and the resulting host toxicity from many antifungal compounds. Cryptococcus neoformans is an opportunistic fungal pathogen of humans that causes life-threatening disease, primarily in immunocompromised patients. Since antifungal therapy for this microorganism is limited, many investigators have explored novel drug targets aim at virulence factors, such as the ability to grow at mammalian physiological temperature (37 degrees C). To address this issue, we used the Agrobacterium tumefaciens gene delivery system to create a random insertion mutagenesis library that was screened for altered growth at elevated temperatures. Among several mutants unable to grow at 37 degrees C, we explored one bearing an interruption in the URA4 gene. This gene encodes dihydroorotase (DHOase) that is involved in the de nova synthesis of pyrimidine ribonucleotides. Loss of the C. neoformans Ura4 protein, by targeted gene interruption, resulted in an expected uracil/uridine auxotrophy and an unexpected high temperature growth defect. in addition, the ura4 mutant displayed phenotypic defects in other prominent virulence factors (melanin, capsule and phospholipase) and reduced stress response compared to wild type and reconstituted strains. Accordingly, this mutant had a decreased survival rate in macrophages and attenuated virulence in a murine model of cryptococcal infection. Quantitative PCR analysis suggests that this biosynthetic pathway is induced during the transition from 30 degrees C to 37 degrees C, and that transcriptional regulation of de nova and salvage pyrimidine pathway are under the control of the Ura4 protein. (C) 2014 the Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-SA license (http://creativecommons.org/licenses/by-nc-sa/3.0/).Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)NIHUniversidade Federal de São Paulo, Dept Ciencias Biol, BR-09972270 Diadema, SP, BrazilUniv Brasilia, Fac Ceilandia, Brasilia, DF, BrazilDuke Univ, Sch Med, Dept Med, Durham, NC 27706 USAUniversidade Federal de São Paulo, Dept Ciencias Biol, BR-09972270 Diadema, SP, BrazilFAPESP: 2007/50536-3FAPESP: 2011/50953-9NIH: AI050128NIH: AI074677Web of Science12-23engElsevier B.V.Fungal Genetics and Biologyhttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policyinfo:eu-repo/semantics/openAccessPyrimidine biosynthesisThermal toleranceBasidiomycete yeastThe role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulenceinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlereponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000341617000003.pdfapplication/pdf2015348${dspace.ui.url}/bitstream/11600/38116/1/WOS000341617000003.pdf4e7430a2b2e92686a61c83a71eee0830MD51open accessTEXTWOS000341617000003.pdf.txtWOS000341617000003.pdf.txtExtracted texttext/plain64823${dspace.ui.url}/bitstream/11600/38116/9/WOS000341617000003.pdf.txt9acf26c491419d196f7972947de48300MD59open accessTHUMBNAILWOS000341617000003.pdf.jpgWOS000341617000003.pdf.jpgIM Thumbnailimage/jpeg7378${dspace.ui.url}/bitstream/11600/38116/11/WOS000341617000003.pdf.jpg190024c14a555cd7d51bfb7f40c2e5caMD511open access11600/381162023-06-05 19:20:16.48open accessoai:repositorio.unifesp.br:11600/38116Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-06-05T22:20:16Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence |
title |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence |
spellingShingle |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence Gontijo, Fabiano Assis de [UNIFESP] Pyrimidine biosynthesis Thermal tolerance Basidiomycete yeast |
title_short |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence |
title_full |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence |
title_fullStr |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence |
title_full_unstemmed |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence |
title_sort |
The role of the de novo pyrimidine biosynthetic pathway in Cryptococcus neoformans high temperature growth and virulence |
author |
Gontijo, Fabiano Assis de [UNIFESP] |
author_facet |
Gontijo, Fabiano Assis de [UNIFESP] Pascon, Renata Castiglioni [UNIFESP] Fernandes, Larissa Machado Junior, Joel [UNIFESP] Alspaugh, J. Andrew Vallim, Marcelo Afonso [UNIFESP] |
author_role |
author |
author2 |
Pascon, Renata Castiglioni [UNIFESP] Fernandes, Larissa Machado Junior, Joel [UNIFESP] Alspaugh, J. Andrew Vallim, Marcelo Afonso [UNIFESP] |
author2_role |
author author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) Universidade de Brasília (UnB) Duke Univ |
dc.contributor.author.fl_str_mv |
Gontijo, Fabiano Assis de [UNIFESP] Pascon, Renata Castiglioni [UNIFESP] Fernandes, Larissa Machado Junior, Joel [UNIFESP] Alspaugh, J. Andrew Vallim, Marcelo Afonso [UNIFESP] |
dc.subject.eng.fl_str_mv |
Pyrimidine biosynthesis Thermal tolerance Basidiomycete yeast |
topic |
Pyrimidine biosynthesis Thermal tolerance Basidiomycete yeast |
description |
Fungal infections are often difficult to treat due to the inherent similarities between fungal and animal cells and the resulting host toxicity from many antifungal compounds. Cryptococcus neoformans is an opportunistic fungal pathogen of humans that causes life-threatening disease, primarily in immunocompromised patients. Since antifungal therapy for this microorganism is limited, many investigators have explored novel drug targets aim at virulence factors, such as the ability to grow at mammalian physiological temperature (37 degrees C). To address this issue, we used the Agrobacterium tumefaciens gene delivery system to create a random insertion mutagenesis library that was screened for altered growth at elevated temperatures. Among several mutants unable to grow at 37 degrees C, we explored one bearing an interruption in the URA4 gene. This gene encodes dihydroorotase (DHOase) that is involved in the de nova synthesis of pyrimidine ribonucleotides. Loss of the C. neoformans Ura4 protein, by targeted gene interruption, resulted in an expected uracil/uridine auxotrophy and an unexpected high temperature growth defect. in addition, the ura4 mutant displayed phenotypic defects in other prominent virulence factors (melanin, capsule and phospholipase) and reduced stress response compared to wild type and reconstituted strains. Accordingly, this mutant had a decreased survival rate in macrophages and attenuated virulence in a murine model of cryptococcal infection. Quantitative PCR analysis suggests that this biosynthetic pathway is induced during the transition from 30 degrees C to 37 degrees C, and that transcriptional regulation of de nova and salvage pyrimidine pathway are under the control of the Ura4 protein. (C) 2014 the Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-SA license (http://creativecommons.org/licenses/by-nc-sa/3.0/). |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-09-01 |
dc.date.accessioned.fl_str_mv |
2016-01-24T14:37:45Z |
dc.date.available.fl_str_mv |
2016-01-24T14:37:45Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
Fungal Genetics and Biology. San Diego: Academic Press Inc Elsevier Science, v. 70, p. 12-23, 2014. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/38116 http://dx.doi.org/10.1016/j.fgb.2014.06.003 |
dc.identifier.issn.none.fl_str_mv |
1087-1845 |
dc.identifier.file.none.fl_str_mv |
WOS000341617000003.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1016/j.fgb.2014.06.003 |
dc.identifier.wos.none.fl_str_mv |
WOS:000341617000003 |
identifier_str_mv |
Fungal Genetics and Biology. San Diego: Academic Press Inc Elsevier Science, v. 70, p. 12-23, 2014. 1087-1845 WOS000341617000003.pdf 10.1016/j.fgb.2014.06.003 WOS:000341617000003 |
url |
http://repositorio.unifesp.br/handle/11600/38116 http://dx.doi.org/10.1016/j.fgb.2014.06.003 |
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eng |
language |
eng |
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Fungal Genetics and Biology |
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http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy info:eu-repo/semantics/openAccess |
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http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy |
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openAccess |
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12-23 |
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Elsevier B.V. |
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Elsevier B.V. |
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