Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas
Autor(a) principal: | |
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Data de Publicação: | 2010 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
dARK ID: | ark:/38995/00130000052h8 |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tde/1563 |
Resumo: | Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis (PCM), a human systemic mycosis, prevalent in Latin America. Extracellular matrix (ECM) is a complex net where collagens, laminin and fibronectin can be found and, when exposed, is the first site for the fungus adhesion. Our aim was to study genes involved in the adhesion process using Representational Difference Analysis (RDA). RDA is a PCR-coupled subtractive method that allows the isolation of genes differentially expressed in two different cDNA populations. Hence, cDNAs were synthesized from RNAs extracted from P. brasiliensis yeast cells adhered to collagen and fibronectin to identify overexpressed genes. Genes involved in a wide range of cellular process were found and PbCtr3 (cooper transporter) and enolase (PbEno) were chosen to further studies. A synthetic peptide (PbCTR3) and the recombinant enolase (rPbEno) were utilized together with the anti-rPbEno polyclonal antibody in functional analysis with ECM components and plasminogen. The studies suggest that P. brasiliensis enolase, in the surface, is able to generate plasmin from plasminogen by plasminogen activator. Therefore, it was also demonstrated that this protein is secreted and able to promote fungus adhesion and invasion to cells. These findings clearly establish the role of enolase in the patogenicity of P. brasiliensis. |
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SOARES, Célia Maria de Almeidahttp://lattes.cnpq.br/8539946335852637http://lattes.cnpq.br/6616012647720101NOGUEIRA, Sarah Veloso2014-07-29T15:26:19Z2010-09-092010-03-11NOGUEIRA, Sarah Veloso. Transcriptional analysis of the accession process by Paracoccidioides brasiliensis and functional characterization of adhesins. 2010. 132 f. Tese (Doutorado em Ciências da Saúde) - Universidade Federal de Goiás, Goiânia, 2010.http://repositorio.bc.ufg.br/tede/handle/tde/1563ark:/38995/00130000052h8Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis (PCM), a human systemic mycosis, prevalent in Latin America. Extracellular matrix (ECM) is a complex net where collagens, laminin and fibronectin can be found and, when exposed, is the first site for the fungus adhesion. Our aim was to study genes involved in the adhesion process using Representational Difference Analysis (RDA). RDA is a PCR-coupled subtractive method that allows the isolation of genes differentially expressed in two different cDNA populations. Hence, cDNAs were synthesized from RNAs extracted from P. brasiliensis yeast cells adhered to collagen and fibronectin to identify overexpressed genes. Genes involved in a wide range of cellular process were found and PbCtr3 (cooper transporter) and enolase (PbEno) were chosen to further studies. A synthetic peptide (PbCTR3) and the recombinant enolase (rPbEno) were utilized together with the anti-rPbEno polyclonal antibody in functional analysis with ECM components and plasminogen. The studies suggest that P. brasiliensis enolase, in the surface, is able to generate plasmin from plasminogen by plasminogen activator. Therefore, it was also demonstrated that this protein is secreted and able to promote fungus adhesion and invasion to cells. These findings clearly establish the role of enolase in the patogenicity of P. brasiliensis.Paraccidioides brasiliensis é o agente etiológico da paracoccidioidomicose (PCM), uma micose sistêmica, prevalente na América Latina. A matriz extracelular (MEC) é uma rede complexa formada por colágeno, laminina, fibronectina, entre outros componentes, que, quando exposta, é o local inicial de adesão do fungo. Nosso objetivo foi estudar genes envolvidos nesse processo de adesão utilizando Análise Diferencial Representacional (RDA). RDA é um método de subtração acoplado a PCR que permite o isolamento de genes diferencialmente expressos entre duas populações de cDNAs diferentes. Assim, cDNAs foram sintetizados a partir de RNAs extraídos de células leveduriformes de P. brasiliensis aderidos à colágeno e fibronectina para identificar genes super-expressos nestas condições. Genes envolvidos com vários processos celulares foram observados e PbCtr3 (transportador de cobre) e enolase (PbEno) foram escolhidos para análises adicionais. Um peptídeo sintético (PbCTR3) e a proteína recombinante (rPbEno) foram utilizados, juntamente com o anticorpo policlonal antirPbEno em análises funcionais com componentes da MEC e plasminogênio. Os estudos sugerem que a enolase de P. brasiliensis, localizada na parede celular, é capaz de gerar plasmina a partir do plasminogênio mediada pelo ativador de plasminôgenio. Além disso, foi também demonstrado que esta proteína é secretada sendo capaz de promover a adesão e invasão do fungo a células. Esses estudos claramente estabelecem o papel da enolase na patogenicidade de P. brasiliensis.Made available in DSpace on 2014-07-29T15:26:19Z (GMT). No. of bitstreams: 1 Tese Sarah Veloso Nogueira.pdf: 4968384 bytes, checksum: 1a3070a4c2260fba10919cd4f6ccd4f4 (MD5) Previous issue date: 2010-03-11application/pdfhttp://repositorio.bc.ufg.br/TEDE/retrieve/4243/Tese%20Sarah%20Veloso%20Nogueira.pdf.jpgporUniversidade Federal de GoiásDoutorado em Medicina TropicalUFGBRCiências da SaúdeParacoccidioides brasiliensisadesinasenolase1.Paracoccidioides brasiliensis 2.EnolaseParacoccidioides brasiliensisadhesinsenolaseCNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIAAnálises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinasTranscriptional analysis of the accession process by Paracoccidioides brasiliensis and functional characterization of adhesinsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGORIGINALTese Sarah Veloso Nogueira.pdfapplication/pdf4968384http://repositorio.bc.ufg.br/tede/bitstreams/2fcd6160-ff9e-4f46-afbc-7c5f968bc328/download1a3070a4c2260fba10919cd4f6ccd4f4MD51TEXTTese Sarah Veloso Nogueira.pdf.txtTese Sarah Veloso Nogueira.pdf.txtExtracted Texttext/plain200588http://repositorio.bc.ufg.br/tede/bitstreams/206da155-8989-4087-9cb8-47e44e61146b/downloade1d8e6c3f4c44576a093edd4da36a664MD52THUMBNAILTese Sarah Veloso Nogueira.pdf.jpgTese Sarah Veloso Nogueira.pdf.jpgGenerated Thumbnailimage/jpeg5590http://repositorio.bc.ufg.br/tede/bitstreams/4e1c59cd-2461-4a26-a023-ff02eb2adaad/downloadb6c23de4aebf6fc75c78a720257036caMD53tde/15632014-07-30 03:12:34.522open.accessoai:repositorio.bc.ufg.br:tde/1563http://repositorio.bc.ufg.br/tedeRepositório InstitucionalPUBhttp://repositorio.bc.ufg.br/oai/requesttasesdissertacoes.bc@ufg.bropendoar:2014-07-30T06:12:34Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)false |
dc.title.por.fl_str_mv |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas |
dc.title.alternative.eng.fl_str_mv |
Transcriptional analysis of the accession process by Paracoccidioides brasiliensis and functional characterization of adhesins |
title |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas |
spellingShingle |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas NOGUEIRA, Sarah Veloso Paracoccidioides brasiliensis adesinas enolase 1.Paracoccidioides brasiliensis 2.Enolase Paracoccidioides brasiliensis adhesins enolase CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
title_short |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas |
title_full |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas |
title_fullStr |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas |
title_full_unstemmed |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas |
title_sort |
Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas |
author |
NOGUEIRA, Sarah Veloso |
author_facet |
NOGUEIRA, Sarah Veloso |
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.authorLattes.fl_str_mv |
http://lattes.cnpq.br/6616012647720101 |
dc.contributor.author.fl_str_mv |
NOGUEIRA, Sarah Veloso |
contributor_str_mv |
SOARES, Célia Maria de Almeida |
dc.subject.por.fl_str_mv |
Paracoccidioides brasiliensis adesinas enolase 1.Paracoccidioides brasiliensis 2.Enolase |
topic |
Paracoccidioides brasiliensis adesinas enolase 1.Paracoccidioides brasiliensis 2.Enolase Paracoccidioides brasiliensis adhesins enolase CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
dc.subject.eng.fl_str_mv |
Paracoccidioides brasiliensis adhesins enolase |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
description |
Paracoccidioides brasiliensis is the causative agent of paracoccidioidomycosis (PCM), a human systemic mycosis, prevalent in Latin America. Extracellular matrix (ECM) is a complex net where collagens, laminin and fibronectin can be found and, when exposed, is the first site for the fungus adhesion. Our aim was to study genes involved in the adhesion process using Representational Difference Analysis (RDA). RDA is a PCR-coupled subtractive method that allows the isolation of genes differentially expressed in two different cDNA populations. Hence, cDNAs were synthesized from RNAs extracted from P. brasiliensis yeast cells adhered to collagen and fibronectin to identify overexpressed genes. Genes involved in a wide range of cellular process were found and PbCtr3 (cooper transporter) and enolase (PbEno) were chosen to further studies. A synthetic peptide (PbCTR3) and the recombinant enolase (rPbEno) were utilized together with the anti-rPbEno polyclonal antibody in functional analysis with ECM components and plasminogen. The studies suggest that P. brasiliensis enolase, in the surface, is able to generate plasmin from plasminogen by plasminogen activator. Therefore, it was also demonstrated that this protein is secreted and able to promote fungus adhesion and invasion to cells. These findings clearly establish the role of enolase in the patogenicity of P. brasiliensis. |
publishDate |
2010 |
dc.date.available.fl_str_mv |
2010-09-09 |
dc.date.issued.fl_str_mv |
2010-03-11 |
dc.date.accessioned.fl_str_mv |
2014-07-29T15:26:19Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
NOGUEIRA, Sarah Veloso. Transcriptional analysis of the accession process by Paracoccidioides brasiliensis and functional characterization of adhesins. 2010. 132 f. Tese (Doutorado em Ciências da Saúde) - Universidade Federal de Goiás, Goiânia, 2010. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tde/1563 |
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ark:/38995/00130000052h8 |
identifier_str_mv |
NOGUEIRA, Sarah Veloso. Transcriptional analysis of the accession process by Paracoccidioides brasiliensis and functional characterization of adhesins. 2010. 132 f. Tese (Doutorado em Ciências da Saúde) - Universidade Federal de Goiás, Goiânia, 2010. ark:/38995/00130000052h8 |
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http://repositorio.bc.ufg.br/tede/handle/tde/1563 |
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por |
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Universidade Federal de Goiás |
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Doutorado em Medicina Tropical |
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UFG |
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BR |
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Ciências da Saúde |
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Universidade Federal de Goiás |
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