Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Outros Autores: | , , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/30533 http://dx.doi.org/10.1074/jbc.M703097200 |
Resumo: | Trypanosoma cruzi is the protozoan parasite that causes Chagas disease or American trypanosomiasis. Kinetoplastid parasites could be considered as model organisms for studying factors involved in posttranscriptional regulation because they control gene expression almost exclusively at this level. the PUF(Pumilio/FBF1) protein family regulates mRNA stability and translation in eukaryotes, and several members have been identified in trypano-somatids. We used a ribonomic approach to identify the putative target mRNAs associated with TcPUF6, a member of the T. cruzi PUF family. TcPUF6 is expressed in discrete sites in the cytoplasm at various stages of the parasite life cycle and is not associated with the translation machinery. the overexpression of a tandem affinity purification-tagged TcPUF6 protein allowed the identification of associated mRNAs by affinity purification assays and microarray hybridization yielding nine putative target mRNAs. Whole expression analysis of transfected parasites showed that the mRNAs associated with TcPUF6 were down-regulated in populations overexpressing TcPUF6. the association of TcPUF6 with the TcDhh1 helicase in vivo and the cellular co-localization of these proteins in epimastigote forms suggest that TcPUF6 promotes degradation of its associated mRNAs through interaction with RNA degradation complexes. Analysis of the mRNA levels of the putative TcPUF6-regulated genes during the parasite life cycle showed that their transcripts were up-regulated in metacyclic trypomastigotes. in these infective forms no co-localization between TcPUF6 and TcDhh1 was observed. Our results suggest that TcPUF6 regulates the half-lives of its associated transcripts via differential association with mRNA degradation complexes throughout its life cycle. |
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Dallagiovanna, BrunoCorrea, AlejandroProbst, Christian M.Holetz, FabiolaSmircich, PabloAguiar, Alessandra Melo deMansur, FernandaSilva, Claudio Vieira da [UNIFESP]Mortara, Renato Arruda [UNIFESP]Garat, BeatrizBuck, Gregory A.Goldenberg, SamuelKrieger, Marco A.Inst Biol Mol ParanaFundacao Oswaldo CruzFac CienciasUniversidade Federal de São Paulo (UNIFESP)Virginia Commonwealth Univ2016-01-24T13:49:40Z2016-01-24T13:49:40Z2008-03-28Journal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 283, n. 13, p. 8266-8273, 2008.0021-9258http://repositorio.unifesp.br/handle/11600/30533http://dx.doi.org/10.1074/jbc.M70309720010.1074/jbc.M703097200WOS:000254288000024Trypanosoma cruzi is the protozoan parasite that causes Chagas disease or American trypanosomiasis. Kinetoplastid parasites could be considered as model organisms for studying factors involved in posttranscriptional regulation because they control gene expression almost exclusively at this level. the PUF(Pumilio/FBF1) protein family regulates mRNA stability and translation in eukaryotes, and several members have been identified in trypano-somatids. We used a ribonomic approach to identify the putative target mRNAs associated with TcPUF6, a member of the T. cruzi PUF family. TcPUF6 is expressed in discrete sites in the cytoplasm at various stages of the parasite life cycle and is not associated with the translation machinery. the overexpression of a tandem affinity purification-tagged TcPUF6 protein allowed the identification of associated mRNAs by affinity purification assays and microarray hybridization yielding nine putative target mRNAs. Whole expression analysis of transfected parasites showed that the mRNAs associated with TcPUF6 were down-regulated in populations overexpressing TcPUF6. the association of TcPUF6 with the TcDhh1 helicase in vivo and the cellular co-localization of these proteins in epimastigote forms suggest that TcPUF6 promotes degradation of its associated mRNAs through interaction with RNA degradation complexes. Analysis of the mRNA levels of the putative TcPUF6-regulated genes during the parasite life cycle showed that their transcripts were up-regulated in metacyclic trypomastigotes. in these infective forms no co-localization between TcPUF6 and TcDhh1 was observed. Our results suggest that TcPUF6 regulates the half-lives of its associated transcripts via differential association with mRNA degradation complexes throughout its life cycle.Inst Biol Mol Parana, BR-81350010 Curitiba, Parana, BrazilFundacao Oswaldo Cruz, BR-21040900 Rio de Janeiro, BrazilFac Ciencias, Lab Interacc Mol, Montevideo 11400, UruguayUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilVirginia Commonwealth Univ, Ctr Study Biol Complex, Richmond, VA 23284 USAUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilWeb of Science8266-8273engAmer Soc Biochemistry Molecular Biology IncJournal of Biological ChemistryFunctional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruziinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/305332022-02-18 10:14:10.1metadata only accessoai:repositorio.unifesp.br:11600/30533Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652022-02-18T13:14:10Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi |
title |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi |
spellingShingle |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi Dallagiovanna, Bruno |
title_short |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi |
title_full |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi |
title_fullStr |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi |
title_full_unstemmed |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi |
title_sort |
Functional genomic characterization of mRNAs associated with TcPUF6, a pumilio-like protein from Trypanosoma cruzi |
author |
Dallagiovanna, Bruno |
author_facet |
Dallagiovanna, Bruno Correa, Alejandro Probst, Christian M. Holetz, Fabiola Smircich, Pablo Aguiar, Alessandra Melo de Mansur, Fernanda Silva, Claudio Vieira da [UNIFESP] Mortara, Renato Arruda [UNIFESP] Garat, Beatriz Buck, Gregory A. Goldenberg, Samuel Krieger, Marco A. |
author_role |
author |
author2 |
Correa, Alejandro Probst, Christian M. Holetz, Fabiola Smircich, Pablo Aguiar, Alessandra Melo de Mansur, Fernanda Silva, Claudio Vieira da [UNIFESP] Mortara, Renato Arruda [UNIFESP] Garat, Beatriz Buck, Gregory A. Goldenberg, Samuel Krieger, Marco A. |
author2_role |
author author author author author author author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Inst Biol Mol Parana Fundacao Oswaldo Cruz Fac Ciencias Universidade Federal de São Paulo (UNIFESP) Virginia Commonwealth Univ |
dc.contributor.author.fl_str_mv |
Dallagiovanna, Bruno Correa, Alejandro Probst, Christian M. Holetz, Fabiola Smircich, Pablo Aguiar, Alessandra Melo de Mansur, Fernanda Silva, Claudio Vieira da [UNIFESP] Mortara, Renato Arruda [UNIFESP] Garat, Beatriz Buck, Gregory A. Goldenberg, Samuel Krieger, Marco A. |
description |
Trypanosoma cruzi is the protozoan parasite that causes Chagas disease or American trypanosomiasis. Kinetoplastid parasites could be considered as model organisms for studying factors involved in posttranscriptional regulation because they control gene expression almost exclusively at this level. the PUF(Pumilio/FBF1) protein family regulates mRNA stability and translation in eukaryotes, and several members have been identified in trypano-somatids. We used a ribonomic approach to identify the putative target mRNAs associated with TcPUF6, a member of the T. cruzi PUF family. TcPUF6 is expressed in discrete sites in the cytoplasm at various stages of the parasite life cycle and is not associated with the translation machinery. the overexpression of a tandem affinity purification-tagged TcPUF6 protein allowed the identification of associated mRNAs by affinity purification assays and microarray hybridization yielding nine putative target mRNAs. Whole expression analysis of transfected parasites showed that the mRNAs associated with TcPUF6 were down-regulated in populations overexpressing TcPUF6. the association of TcPUF6 with the TcDhh1 helicase in vivo and the cellular co-localization of these proteins in epimastigote forms suggest that TcPUF6 promotes degradation of its associated mRNAs through interaction with RNA degradation complexes. Analysis of the mRNA levels of the putative TcPUF6-regulated genes during the parasite life cycle showed that their transcripts were up-regulated in metacyclic trypomastigotes. in these infective forms no co-localization between TcPUF6 and TcDhh1 was observed. Our results suggest that TcPUF6 regulates the half-lives of its associated transcripts via differential association with mRNA degradation complexes throughout its life cycle. |
publishDate |
2008 |
dc.date.issued.fl_str_mv |
2008-03-28 |
dc.date.accessioned.fl_str_mv |
2016-01-24T13:49:40Z |
dc.date.available.fl_str_mv |
2016-01-24T13:49:40Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Journal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 283, n. 13, p. 8266-8273, 2008. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/30533 http://dx.doi.org/10.1074/jbc.M703097200 |
dc.identifier.issn.none.fl_str_mv |
0021-9258 |
dc.identifier.doi.none.fl_str_mv |
10.1074/jbc.M703097200 |
dc.identifier.wos.none.fl_str_mv |
WOS:000254288000024 |
identifier_str_mv |
Journal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 283, n. 13, p. 8266-8273, 2008. 0021-9258 10.1074/jbc.M703097200 WOS:000254288000024 |
url |
http://repositorio.unifesp.br/handle/11600/30533 http://dx.doi.org/10.1074/jbc.M703097200 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Journal of Biological Chemistry |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
8266-8273 |
dc.publisher.none.fl_str_mv |
Amer Soc Biochemistry Molecular Biology Inc |
publisher.none.fl_str_mv |
Amer Soc Biochemistry Molecular Biology Inc |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
|
_version_ |
1802764108470157312 |