Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp.
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/5811 |
Resumo: | The Paracoccidioides genus comprises a complex of pathogenic fungi that are the etiologic agents of paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin America. The infection begins after inhalation of fungal propagules, which reach the epithelium of the alveoli where the transition from the mycelial to the pathogenic yeast form. Host elevated temperature triggers the morphological switch, which is necessary for fungal pathogenicity. The cAMP/protein kinase A (PKA) signaling pathway has been shown to be important in controlling morphological changes and the pathogenicity of several pathogenic fungi. Evidence also highlights the importance of the cAMP/PKA pathway in the morphological transition of Paracoccidioides. PKA is the major effector of this signaling pathway. The protein is an inactive tetramer composed of regulatory subunit, encoded by the BCY1 gene; and catalytic subunit, encoded by the TPK2 gene. Upon binding of cAMP to the regulatory subunits, the catalytic subunits dissociate and become active. Activated PKA subsequently phosphorylates protein kinases, transcription factors, and other substrates to control several biological processes. In this study, we evaluated the expression and interactions of Tpk2 protein Paracoccididioides spp. The Tpk2 is present in mycelium decreased during the initial stages of transition phases, and increases again at the end of differentiation, with maximal levels in yeast. We analyzed the interactions of recombinant Tpk2p with Paracoccidioides proteins using pull-down assays followed by MS analysis. Two interacting proteins were identified: the heat shock protein 90 (Hsp90) and a conserved hypothetical protein with a MFS domain. Hsp90 is involved in the regulation of morphogenesis, development and virulence in several thermal dimorphic fungi. These data are important for understanding the mechanisms that trigger the transition phases in Paracoccidioides. |
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Furlaneto, Silvia M. Salem Izacchttp://lattes.cnpq.br/5542866107144940Furlaneto, Silvia M. Salem IzaccParente, Juliana AlvesBaeza, Lilian Cristianehttp://lattes.cnpq.br/0149863389258237Teixeira, Mirian Vieira2016-08-04T11:30:40Z2016-03-10TEIXEIRA, M. V. Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. 2016. 62 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2016.http://repositorio.bc.ufg.br/tede/handle/tede/5811ark:/38995/0013000003t73The Paracoccidioides genus comprises a complex of pathogenic fungi that are the etiologic agents of paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin America. The infection begins after inhalation of fungal propagules, which reach the epithelium of the alveoli where the transition from the mycelial to the pathogenic yeast form. Host elevated temperature triggers the morphological switch, which is necessary for fungal pathogenicity. The cAMP/protein kinase A (PKA) signaling pathway has been shown to be important in controlling morphological changes and the pathogenicity of several pathogenic fungi. Evidence also highlights the importance of the cAMP/PKA pathway in the morphological transition of Paracoccidioides. PKA is the major effector of this signaling pathway. The protein is an inactive tetramer composed of regulatory subunit, encoded by the BCY1 gene; and catalytic subunit, encoded by the TPK2 gene. Upon binding of cAMP to the regulatory subunits, the catalytic subunits dissociate and become active. Activated PKA subsequently phosphorylates protein kinases, transcription factors, and other substrates to control several biological processes. In this study, we evaluated the expression and interactions of Tpk2 protein Paracoccididioides spp. The Tpk2 is present in mycelium decreased during the initial stages of transition phases, and increases again at the end of differentiation, with maximal levels in yeast. We analyzed the interactions of recombinant Tpk2p with Paracoccidioides proteins using pull-down assays followed by MS analysis. Two interacting proteins were identified: the heat shock protein 90 (Hsp90) and a conserved hypothetical protein with a MFS domain. Hsp90 is involved in the regulation of morphogenesis, development and virulence in several thermal dimorphic fungi. These data are important for understanding the mechanisms that trigger the transition phases in Paracoccidioides.O gênero Paracoccidioides compreende um complexo de fungos patogênicos, que são os agentes etiológicos da paracoccidioidomicose (PCM), a micose sistêmica mais prevalente na América Latina. A infecção inicia-se com a inalação de propágulos do fungo, que atingem o epitélio dos alvéolos pulmonares, onde ocorre à transição da forma de micélio para a forma patogênica, de levedura. Há evidências de que a temperatura seja o principal fator responsável pela diferenciação celular desses fungos, e sua patogenicidade é frequentemente associada com a transição dimórfica. A via de sinalização cAMP/ proteína quinase A (PKA) controla alterações morfológicas e de virulência/patogenicidade em várias espécies de fungos patogênicos humanos. Evidências apontam também para a importância da via cAMP/PKA em Paracoccidioides spp. A PKA é o principal efetor desta via de sinalização. A proteína na forma inativa é um tetrâmero composto de subunidade regulatória, codificada pelo gene BCY1; e subunidade catalítica, codificada pelo gene TPK2. Após a ligação de cAMP às subunidades regulatórias, as subunidades catalíticas dissociam-se e tornam-se ativas. Ativada a PKA fosforila proteína-quinases, fatores de transcrição, e outros substratos para controlar diversos processos biológicos. Neste estudo, avaliamos a expressão e as interações da proteína Tpk2 de Paracoccididioides spp. A Tpk2 está presente em micélio, diminui nos estágios iniciais da transição de fases e volta a aumentar no final da diferenciação, apresentando níveis máximos na levedura. Foram analisadas as interações de Tpk2p recombinante com proteínas de Paracoccidioides utilizando ensaios de pull-down, seguido por análise de MS. Foram identificadas duas proteínas que interagem: a proteína de choque térmico 90 (Hsp90) e uma proteína hipotética conservada com um domínio MFS. Hsp90 está envolvido na regulação da morfogênese, desenvolvimento e virulência em vários fungos dimórficos térmicos. Estes dados são importantes para entendimento dos mecanismos que disparam a transição de fases em Paracoccidioides spp.Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-04T11:29:03Z No. of bitstreams: 2 Dissertação - Mirian Vieira Teixeira - 2016.pdf: 1440228 bytes, checksum: a429eb9e9ac048fbcc06d59fca852c03 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-04T11:30:39Z (GMT) No. of bitstreams: 2 Dissertação - Mirian Vieira Teixeira - 2016.pdf: 1440228 bytes, checksum: a429eb9e9ac048fbcc06d59fca852c03 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2016-08-04T11:30:40Z (GMT). No. of bitstreams: 2 Dissertação - Mirian Vieira Teixeira - 2016.pdf: 1440228 bytes, checksum: a429eb9e9ac048fbcc06d59fca852c03 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-03-10Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqFundação de Amparo à Pesquisa do Estado de Goiás - FAPEGapplication/pdfporUniversidade Federal de GoiásPrograma de Pós-graduação em Genetica e Biologia MolecularUFGBrasilInstituto de Ciências Biológicas - ICB (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessParacoccidioidesTransição dimórficaVia cAMP/PKAEnsaios de pull downParacoccidioidesDimorphic transitioncAMP/PKA pathwayPull-down assaysGENETICA::GENETICA HUMANA E MEDICAAnálise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp.Analysis of the expression and interactions of the catalytic subunit of PKA in pathogenic fungus Paracoccidioides ssp.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-7235106986317239737600600600600600-38727721178273734044510125209561567543-2555911436985713659-961409807440757778reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. |
dc.title.alternative.eng.fl_str_mv |
Analysis of the expression and interactions of the catalytic subunit of PKA in pathogenic fungus Paracoccidioides ssp. |
title |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. |
spellingShingle |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. Teixeira, Mirian Vieira Paracoccidioides Transição dimórfica Via cAMP/PKA Ensaios de pull down Paracoccidioides Dimorphic transition cAMP/PKA pathway Pull-down assays GENETICA::GENETICA HUMANA E MEDICA |
title_short |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. |
title_full |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. |
title_fullStr |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. |
title_full_unstemmed |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. |
title_sort |
Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. |
author |
Teixeira, Mirian Vieira |
author_facet |
Teixeira, Mirian Vieira |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Furlaneto, Silvia M. Salem Izacc |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5542866107144940 |
dc.contributor.referee1.fl_str_mv |
Furlaneto, Silvia M. Salem Izacc |
dc.contributor.referee2.fl_str_mv |
Parente, Juliana Alves |
dc.contributor.referee3.fl_str_mv |
Baeza, Lilian Cristiane |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/0149863389258237 |
dc.contributor.author.fl_str_mv |
Teixeira, Mirian Vieira |
contributor_str_mv |
Furlaneto, Silvia M. Salem Izacc Furlaneto, Silvia M. Salem Izacc Parente, Juliana Alves Baeza, Lilian Cristiane |
dc.subject.por.fl_str_mv |
Paracoccidioides Transição dimórfica Via cAMP/PKA Ensaios de pull down |
topic |
Paracoccidioides Transição dimórfica Via cAMP/PKA Ensaios de pull down Paracoccidioides Dimorphic transition cAMP/PKA pathway Pull-down assays GENETICA::GENETICA HUMANA E MEDICA |
dc.subject.eng.fl_str_mv |
Paracoccidioides Dimorphic transition cAMP/PKA pathway Pull-down assays |
dc.subject.cnpq.fl_str_mv |
GENETICA::GENETICA HUMANA E MEDICA |
description |
The Paracoccidioides genus comprises a complex of pathogenic fungi that are the etiologic agents of paracoccidioidomycosis (PCM), the most prevalent systemic mycosis in Latin America. The infection begins after inhalation of fungal propagules, which reach the epithelium of the alveoli where the transition from the mycelial to the pathogenic yeast form. Host elevated temperature triggers the morphological switch, which is necessary for fungal pathogenicity. The cAMP/protein kinase A (PKA) signaling pathway has been shown to be important in controlling morphological changes and the pathogenicity of several pathogenic fungi. Evidence also highlights the importance of the cAMP/PKA pathway in the morphological transition of Paracoccidioides. PKA is the major effector of this signaling pathway. The protein is an inactive tetramer composed of regulatory subunit, encoded by the BCY1 gene; and catalytic subunit, encoded by the TPK2 gene. Upon binding of cAMP to the regulatory subunits, the catalytic subunits dissociate and become active. Activated PKA subsequently phosphorylates protein kinases, transcription factors, and other substrates to control several biological processes. In this study, we evaluated the expression and interactions of Tpk2 protein Paracoccididioides spp. The Tpk2 is present in mycelium decreased during the initial stages of transition phases, and increases again at the end of differentiation, with maximal levels in yeast. We analyzed the interactions of recombinant Tpk2p with Paracoccidioides proteins using pull-down assays followed by MS analysis. Two interacting proteins were identified: the heat shock protein 90 (Hsp90) and a conserved hypothetical protein with a MFS domain. Hsp90 is involved in the regulation of morphogenesis, development and virulence in several thermal dimorphic fungi. These data are important for understanding the mechanisms that trigger the transition phases in Paracoccidioides. |
publishDate |
2016 |
dc.date.accessioned.fl_str_mv |
2016-08-04T11:30:40Z |
dc.date.issued.fl_str_mv |
2016-03-10 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
TEIXEIRA, M. V. Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. 2016. 62 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2016. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/5811 |
dc.identifier.dark.fl_str_mv |
ark:/38995/0013000003t73 |
identifier_str_mv |
TEIXEIRA, M. V. Análise da expressão e das interações da subunidade catalítica da PKA do fungo patogênico Paracoccidioides ssp. 2016. 62 f. Dissertação (Mestrado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2016. ark:/38995/0013000003t73 |
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http://repositorio.bc.ufg.br/tede/handle/tede/5811 |
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por |
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por |
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4510125209561567543 |
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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/ |
eu_rights_str_mv |
openAccess |
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Universidade Federal de Goiás |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Genetica e Biologia Molecular |
dc.publisher.initials.fl_str_mv |
UFG |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Instituto de Ciências Biológicas - ICB (RG) |
publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.source.none.fl_str_mv |
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