Análises transcricionais no processo de adesão por Paracoccidioides brasiliensis e caracterização funcional de adesinas

Detalhes bibliográficos
Autor(a) principal: NOGUEIRA, Sarah Veloso
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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spelling 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). 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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
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