Respostas transcricionais de Paracoccidioides ao estresse nitrosativo
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/5721 |
Resumo: | Paracoccidioides is a thermal dimorphic fungus that causes Paracoccidioidomycosis (PCM), an endemic disease in Latin America. Immune cells have a variety of defense mechanisms against pathogens, such as the production and release of nitric oxide (NO), one of the reactive nitrogen species (RNS). RNS react with cellular components, resulting in damage to DNA and membranes, inhibition of repiration and inactivation of cellular enzymes. To understand how Paracoccidioides responds to nitrosative stress, this study aims to identify genes that might contribute to this response. The Paracoccidioides yeast cells grouth was evaluated in the presence of various concentrations of sodium nitrite (NaNO2). The nitrosative stress was confirmed by quantification of nitrite in the cultures supernatant and by the inhibition of the cytochrome c oxidase (complex IV) activity. The transcriptional analyzes were performed by sequencing a cDNA library constructed with Paracoccidioides mRNA obtained after incubation of fungal cells with 500 µM NaNO2 during 1 h. The results show the induction of transcripts related to several cellular pathways, including genes of mitochondrial electron transport chain. The expression of selected transcripts was confirmed by quantitative real time RT-PCR. Moreover, mitochondrial activity measured by XTT method was reduced in the presence of NaNO2 during the first 2 hours of treatment. Additionally, the NADP+/NADPH ratio is lower during nitrosative stress, as demonstrated. The results suggest that the induction of transcripts associated with energy production during nitrosative stress may reflect a compensatory effect on the inhibition of enzymes that carry out this metabolic function, probably to ensure the production of energy and/or contribute to the redox balance by generation of NADPH. |
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Soares, Célia Maria de Almeidahttp://lattes.cnpq.br/8539946335852637Soares, Célia Maria de AlmeidaPereira, MaristelaParente, Juliana Alveshttp://lattes.cnpq.br/8785505426877057Naves, Priscila Elias Campos2016-07-14T13:01:06Z2012-08-30NAVES, P. E. C. Respostas transcricionais de Paracoccidioides ao estresse nitrosativo. 2012. 70 f. Dissertação (Mestrado em Biologia) - Universidade Federal de Goiás, Goiânia, 2012.http://repositorio.bc.ufg.br/tede/handle/tede/5721ark:/38995/0013000009qt6Paracoccidioides is a thermal dimorphic fungus that causes Paracoccidioidomycosis (PCM), an endemic disease in Latin America. Immune cells have a variety of defense mechanisms against pathogens, such as the production and release of nitric oxide (NO), one of the reactive nitrogen species (RNS). RNS react with cellular components, resulting in damage to DNA and membranes, inhibition of repiration and inactivation of cellular enzymes. To understand how Paracoccidioides responds to nitrosative stress, this study aims to identify genes that might contribute to this response. The Paracoccidioides yeast cells grouth was evaluated in the presence of various concentrations of sodium nitrite (NaNO2). The nitrosative stress was confirmed by quantification of nitrite in the cultures supernatant and by the inhibition of the cytochrome c oxidase (complex IV) activity. The transcriptional analyzes were performed by sequencing a cDNA library constructed with Paracoccidioides mRNA obtained after incubation of fungal cells with 500 µM NaNO2 during 1 h. The results show the induction of transcripts related to several cellular pathways, including genes of mitochondrial electron transport chain. The expression of selected transcripts was confirmed by quantitative real time RT-PCR. Moreover, mitochondrial activity measured by XTT method was reduced in the presence of NaNO2 during the first 2 hours of treatment. Additionally, the NADP+/NADPH ratio is lower during nitrosative stress, as demonstrated. The results suggest that the induction of transcripts associated with energy production during nitrosative stress may reflect a compensatory effect on the inhibition of enzymes that carry out this metabolic function, probably to ensure the production of energy and/or contribute to the redox balance by generation of NADPH.Paracoccidioides é um fungo termodimórfico causador da Paracoccidioidomicose (PCM), uma doença endêmica na América Latina. Células do sistema imune humano possuem uma variedade de mecanismos de defesa contra patógenos, como a produção e liberação de óxido nítrico (NO), uma das espécies reativas de nitrogênio (RNS). RNS reagem com os componentes celulares, o que resulta em danos ao DNA e membranas, inibição da respiração e inativação de enzimas celulares. Para entender como Paracoccidioides responde ao estresse nitrosativo, esse trabalho tem como objetivo identificar genes que possam contribuir para esta resposta. O crescimento de células de levedura de Paracoccidioides foi avaliado na presença de várias concentrações de nitrito de sódio (NaNO2). O estresse nitrosativo foi confirmado através da quantificação de nitrito no sobrenadante das culturas e da inibição da atividade da enzima citocromo c oxidase (Complexo IV). As análises transcricionais foram feitas a partir do sequenciamento da biblioteca de cDNA construída utilizando-se mRNA obtido após incubação de células leveduriformes de Paracoccidioides na presença de 500µM de NaNO2 durante 1 hora. Os resultados mostram a indução de transcritos relacionados a várias vias celulares, incluindo genes da cadeia mitocondrial transportadora de elétrons. A expressão de alguns desses transcritos foi confirmada por PCR quantitativo em tempo real. Adicionalmente, a atividade mitocondrial, avaliada através do método do XTT, apresentou-se reduzida na presença de NaNO2 durante as primeiras 2 horas de tratamento. A razão NADP+/NADPH foi avaliada, apresentando-se menor durante o estresse nitrosativo. Os resultados sugerem que a indução de transcritos relacionados à produção de energia durante o estresse nitrosativo possa refletir um efeito compensatório sobre a inibição de enzimas que desempenham esta função metabólica, provavelmente para assegurar a produção de energia e/ou contribuir para o balanço redox através da geração de NADPH.Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-07-13T21:03:27Z No. of bitstreams: 2 Dissertação - Priscila Elias Campos Naves - 2012.pdf: 1687485 bytes, checksum: 4d21dacd2ba42ae2bd0c8df7e82075b0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-07-14T13:01:06Z (GMT) No. of bitstreams: 2 Dissertação - Priscila Elias Campos Naves - 2012.pdf: 1687485 bytes, checksum: 4d21dacd2ba42ae2bd0c8df7e82075b0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2016-07-14T13:01:06Z (GMT). No. of bitstreams: 2 Dissertação - Priscila Elias Campos Naves - 2012.pdf: 1687485 bytes, checksum: 4d21dacd2ba42ae2bd0c8df7e82075b0 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2012-08-30Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfporUniversidade Federal de GoiásPrograma de Pós-graduação em Biologia (ICB)UFGBrasilInstituto de Ciências Biológicas - ICB (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessParacoccidioidesEstresse nitrosativoTranscritomaParacoccidioidesNitrosative stressTranscriptomeBIOQUIMICA::BIOLOGIA MOLECULARRespostas transcricionais de Paracoccidioides ao estresse nitrosativoTranscriptional responses of Paracoccidioides to nitrosative stressinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis6883982777473437920600600600600-387277211782737340439621439903280520722075167498588264571reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo |
dc.title.alternative.eng.fl_str_mv |
Transcriptional responses of Paracoccidioides to nitrosative stress |
title |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo |
spellingShingle |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo Naves, Priscila Elias Campos Paracoccidioides Estresse nitrosativo Transcritoma Paracoccidioides Nitrosative stress Transcriptome BIOQUIMICA::BIOLOGIA MOLECULAR |
title_short |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo |
title_full |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo |
title_fullStr |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo |
title_full_unstemmed |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo |
title_sort |
Respostas transcricionais de Paracoccidioides ao estresse nitrosativo |
author |
Naves, Priscila Elias Campos |
author_facet |
Naves, Priscila Elias Campos |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Soares, Célia Maria de Almeida |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/8539946335852637 |
dc.contributor.referee1.fl_str_mv |
Soares, Célia Maria de Almeida |
dc.contributor.referee2.fl_str_mv |
Pereira, Maristela |
dc.contributor.referee3.fl_str_mv |
Parente, Juliana Alves |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/8785505426877057 |
dc.contributor.author.fl_str_mv |
Naves, Priscila Elias Campos |
contributor_str_mv |
Soares, Célia Maria de Almeida Soares, Célia Maria de Almeida Pereira, Maristela Parente, Juliana Alves |
dc.subject.por.fl_str_mv |
Paracoccidioides Estresse nitrosativo Transcritoma |
topic |
Paracoccidioides Estresse nitrosativo Transcritoma Paracoccidioides Nitrosative stress Transcriptome BIOQUIMICA::BIOLOGIA MOLECULAR |
dc.subject.eng.fl_str_mv |
Paracoccidioides Nitrosative stress Transcriptome |
dc.subject.cnpq.fl_str_mv |
BIOQUIMICA::BIOLOGIA MOLECULAR |
description |
Paracoccidioides is a thermal dimorphic fungus that causes Paracoccidioidomycosis (PCM), an endemic disease in Latin America. Immune cells have a variety of defense mechanisms against pathogens, such as the production and release of nitric oxide (NO), one of the reactive nitrogen species (RNS). RNS react with cellular components, resulting in damage to DNA and membranes, inhibition of repiration and inactivation of cellular enzymes. To understand how Paracoccidioides responds to nitrosative stress, this study aims to identify genes that might contribute to this response. The Paracoccidioides yeast cells grouth was evaluated in the presence of various concentrations of sodium nitrite (NaNO2). The nitrosative stress was confirmed by quantification of nitrite in the cultures supernatant and by the inhibition of the cytochrome c oxidase (complex IV) activity. The transcriptional analyzes were performed by sequencing a cDNA library constructed with Paracoccidioides mRNA obtained after incubation of fungal cells with 500 µM NaNO2 during 1 h. The results show the induction of transcripts related to several cellular pathways, including genes of mitochondrial electron transport chain. The expression of selected transcripts was confirmed by quantitative real time RT-PCR. Moreover, mitochondrial activity measured by XTT method was reduced in the presence of NaNO2 during the first 2 hours of treatment. Additionally, the NADP+/NADPH ratio is lower during nitrosative stress, as demonstrated. The results suggest that the induction of transcripts associated with energy production during nitrosative stress may reflect a compensatory effect on the inhibition of enzymes that carry out this metabolic function, probably to ensure the production of energy and/or contribute to the redox balance by generation of NADPH. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012-08-30 |
dc.date.accessioned.fl_str_mv |
2016-07-14T13:01:06Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
NAVES, P. E. C. Respostas transcricionais de Paracoccidioides ao estresse nitrosativo. 2012. 70 f. Dissertação (Mestrado em Biologia) - Universidade Federal de Goiás, Goiânia, 2012. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/5721 |
dc.identifier.dark.fl_str_mv |
ark:/38995/0013000009qt6 |
identifier_str_mv |
NAVES, P. E. C. Respostas transcricionais de Paracoccidioides ao estresse nitrosativo. 2012. 70 f. Dissertação (Mestrado em Biologia) - Universidade Federal de Goiás, Goiânia, 2012. ark:/38995/0013000009qt6 |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/5721 |
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por |
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por |
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3962143990328052072 |
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http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
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http://creativecommons.org/licenses/by-nc-nd/4.0/ |
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Universidade Federal de Goiás |
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UFG |
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Universidade Federal de Goiás |
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