The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.14/9267 |
Resumo: | Chlamydia trachomatis is the most common cause of sexually transmitted bacterial infection in Europe and in the United States. It is a Gram negative obligate intracellular pathogen with a unique biphasic development cycle, possessing two distinct morphological forms, the elementary body (EB) and the reticulate body (RB). The EBs are the infectious form and are able to attach and invade mammalian cells. This process is likely dependent on the Type III Secretion System, which allows secretion of Chlamydia effectors that modulate the actin skeleton of the host cell and promote invasion. One of the first effectors to be secreted is TarP (Translocated Actin Recruiting Protein). Orthologs of this effector are present in all Chlamydia with some polymorphisms associated with some subdomains. An example is the tyrosine rich domain, which is only present in the TarP homolog of the C. trachomatis species. Despite this difference, all chlamydial TarP are able to recruit and nucleate actin to promote the internalisation of Chlamydia. Since early secretion of TarP is crucial for invasion it was hypothesized that this secretion, in order to be efficient must be facilitated by a Type III Secretion chaperone. These usually are small proteins (13-16 kDa) with an acidic pI and a secondary structure motif of α-β-β-β-α-β-β. Three putative Chlamydia trachomatis Type III secretion chaperones, CT043 (Slc1), CT663 (Slc2), CT088 (Scc1), which are present in EBs have been predicted by their secondary structures. The interaction of these chaperones with the N-terminal domain of TarP (amino acids 1-200) was tested using pull-down assays. A specific interaction between TarP and Slc1 was observed, but not with Slc2 or Scc1. Using a bacterial two-hybrid assay, we evaluated the effects of mutating TarP(1-100) on its interaction with Slc1. The objective was to identify critical amino acid residues involved in chaperone binding. Using several mutants TarP derivatives, we were able to conclude that the interaction between TarP and Slc1 may depend of both specific amino acids and hydrophobic interactions. |
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The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043Chlamydia trachomatis is the most common cause of sexually transmitted bacterial infection in Europe and in the United States. It is a Gram negative obligate intracellular pathogen with a unique biphasic development cycle, possessing two distinct morphological forms, the elementary body (EB) and the reticulate body (RB). The EBs are the infectious form and are able to attach and invade mammalian cells. This process is likely dependent on the Type III Secretion System, which allows secretion of Chlamydia effectors that modulate the actin skeleton of the host cell and promote invasion. One of the first effectors to be secreted is TarP (Translocated Actin Recruiting Protein). Orthologs of this effector are present in all Chlamydia with some polymorphisms associated with some subdomains. An example is the tyrosine rich domain, which is only present in the TarP homolog of the C. trachomatis species. Despite this difference, all chlamydial TarP are able to recruit and nucleate actin to promote the internalisation of Chlamydia. Since early secretion of TarP is crucial for invasion it was hypothesized that this secretion, in order to be efficient must be facilitated by a Type III Secretion chaperone. These usually are small proteins (13-16 kDa) with an acidic pI and a secondary structure motif of α-β-β-β-α-β-β. Three putative Chlamydia trachomatis Type III secretion chaperones, CT043 (Slc1), CT663 (Slc2), CT088 (Scc1), which are present in EBs have been predicted by their secondary structures. The interaction of these chaperones with the N-terminal domain of TarP (amino acids 1-200) was tested using pull-down assays. A specific interaction between TarP and Slc1 was observed, but not with Slc2 or Scc1. Using a bacterial two-hybrid assay, we evaluated the effects of mutating TarP(1-100) on its interaction with Slc1. The objective was to identify critical amino acid residues involved in chaperone binding. Using several mutants TarP derivatives, we were able to conclude that the interaction between TarP and Slc1 may depend of both specific amino acids and hydrophobic interactions.A Chlamydia trachomatis é responsável por uma das infecções bacterianas sexualmente transmissíveis mais comuns na Europa e nos EUA. Esta bactéria Gram negativa é um patogénico intracelular obrigatório, com um ciclo biológico bifásico, no qual se observam duas morfologias bacterianas diferentes, o Corpo Elementar (CE) e o Corpo Reticular (CR). As formas infectantes desta bactéria são os CEs que têm capacidade de se agregar e invadir as células dos mamíferos. Este processo parece ser dependente do Sistema de Secreção Tipo III (T3SS), permitindo a secreção de efectores de Chlamydia para as células do hospedeiro, os quais modulam o esqueleto de actina das células eucariótas promovendo a invasão. Um dos primeiros efectores secretados pela Chlamydia designa-se TarP (Translocated Actin Recruiting Protein). Efectores ortólogos ao TarP estão presentes em todas as espécies do género Chlamydia. Estes ortólogos apresentam polimorfismos associados com alguns subdomínios, como é o caso o domínio rico em tirosina, que apenas se encontra presente no TarP descrito para C. trachomatis. Apesar das diferenças verificadas nos ortólogos das diferentes espécies de Chlamydia, todos têm a capacidade de recrutar e agregar actina permitindo a internalização da bactéria por parte da célula. Uma vez que, numa fase inicial da invasão, a secreção do TarP parece ser indispensável para a mesma, poderia pensar-se que, para que esta secreção seja eficiente, deve ser facilitada por uma chaperona do T3SS. Estas chaperonas são, normalmente, proteínas de baixo peso molecular (13-16 kDa), com um ponto isoeléctrico acídico e uma estrutura secundária com um motivo α-β-β-β-α-β-β. Através do estudo da estrutura secundária previram-se três chaperonas do T3SS, CT043 (Slc1), CT663 (Slc2) CT088 (Scc1), presentes em CEs de Chlamydia. A interacção destas chaperonas com o domínio N-terminal do TarP (aa 1-200) testou-se através de ensaios de co-imunoprecipitação. Observou-se que, das três chaperonas testadas, o TarP unicamente interage de forma específica com a chaperona Slc1. Tendo como base estes resultados, utilizou-se um ensaio de duplo híbrido para avaliar quais os efeitos que mutações a nível do N-ternimal do TarP (1-100) têm na sua interacção com Slc1. Este estudo teve como objectivo identificar os aminoácidos que estão envolvidos na ligação do TarP à chaperona Slc1. Através da utilização de diversos mutantes no N-terminal do TarP, concluiu-se que a interacção entre o TarP e Slc1 parece depender tanto de aminoácidos específicos como de interacções hidrofóbicas entre TarP e Slc1.Carabeo, ReyVeritati - Repositório Institucional da Universidade Católica PortuguesaPedrosa, António José Santos Tedim Sousa2012-10-16T08:50:54Z2011-0720112011-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10400.14/9267enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-08-01T23:50:07Zoai:repositorio.ucp.pt:10400.14/9267Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:08:19.576107Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 |
title |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 |
spellingShingle |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 Pedrosa, António José Santos Tedim Sousa |
title_short |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 |
title_full |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 |
title_fullStr |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 |
title_full_unstemmed |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 |
title_sort |
The characterization of the possible interaction between the Chlamydia effector tarp and its outative chaperone CT043 |
author |
Pedrosa, António José Santos Tedim Sousa |
author_facet |
Pedrosa, António José Santos Tedim Sousa |
author_role |
author |
dc.contributor.none.fl_str_mv |
Carabeo, Rey Veritati - Repositório Institucional da Universidade Católica Portuguesa |
dc.contributor.author.fl_str_mv |
Pedrosa, António José Santos Tedim Sousa |
description |
Chlamydia trachomatis is the most common cause of sexually transmitted bacterial infection in Europe and in the United States. It is a Gram negative obligate intracellular pathogen with a unique biphasic development cycle, possessing two distinct morphological forms, the elementary body (EB) and the reticulate body (RB). The EBs are the infectious form and are able to attach and invade mammalian cells. This process is likely dependent on the Type III Secretion System, which allows secretion of Chlamydia effectors that modulate the actin skeleton of the host cell and promote invasion. One of the first effectors to be secreted is TarP (Translocated Actin Recruiting Protein). Orthologs of this effector are present in all Chlamydia with some polymorphisms associated with some subdomains. An example is the tyrosine rich domain, which is only present in the TarP homolog of the C. trachomatis species. Despite this difference, all chlamydial TarP are able to recruit and nucleate actin to promote the internalisation of Chlamydia. Since early secretion of TarP is crucial for invasion it was hypothesized that this secretion, in order to be efficient must be facilitated by a Type III Secretion chaperone. These usually are small proteins (13-16 kDa) with an acidic pI and a secondary structure motif of α-β-β-β-α-β-β. Three putative Chlamydia trachomatis Type III secretion chaperones, CT043 (Slc1), CT663 (Slc2), CT088 (Scc1), which are present in EBs have been predicted by their secondary structures. The interaction of these chaperones with the N-terminal domain of TarP (amino acids 1-200) was tested using pull-down assays. A specific interaction between TarP and Slc1 was observed, but not with Slc2 or Scc1. Using a bacterial two-hybrid assay, we evaluated the effects of mutating TarP(1-100) on its interaction with Slc1. The objective was to identify critical amino acid residues involved in chaperone binding. Using several mutants TarP derivatives, we were able to conclude that the interaction between TarP and Slc1 may depend of both specific amino acids and hydrophobic interactions. |
publishDate |
2011 |
dc.date.none.fl_str_mv |
2011-07 2011 2011-07-01T00:00:00Z 2012-10-16T08:50:54Z |
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.uri.fl_str_mv |
http://hdl.handle.net/10400.14/9267 |
url |
http://hdl.handle.net/10400.14/9267 |
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eng |
language |
eng |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799131752205123584 |