Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi

Autores
Westergaard, Gaston G.; Bercovich, Natalia; Reinert, Marina Daniela; Vazquez, Martin Pablo
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
There are only a few reported nuclear localization signals (NLS) in trypanosomes despite intensive research on nuclear metabolic processes such as mRNA processing and transcription during the recent past. Moreover, there are only two reports for a monopartite (La protein) and bipartite (H2B histone, ESAG8) NLS in Trypanosoma brucei. We decided to investigate a NLS in Trypanosoma cruzi by selecting p14, a small RNA recognition motif (RRM) containing protein involved in the splicing process in the nucleus. Its small size (117 amino acids), and an optimized streamlined workflow for analysis in T. cruzi, allowed us to define a region of basic amino acids (RRKRRR) located at the C-terminus that is necessary for nuclear localization. However, the NLS for p14 appeared to be more complex since the signature RRKRRR alone is necessary but not sufficient to direct heterologous proteins, such as GFP, to the nucleus. Since p14 interacts strongly with splicing factor SF3b155, a much larger protein, we designed a p14 variant unable to interact with it. The results allowed us to discard the notion that p14 is entering the nucleus, or is retained within, as the sole consequence of being part of a larger complex. Extensive mapping showed that all of the information for nuclear import resides within the small p14 protein in a bipartite NLS composed of the signature RRKRRR and a region of the RRM domain. Thus, NLS definition in T. cruzi is more complex than previously described.
Fil: Westergaard, Gaston G.. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
Fil: Bercovich, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Max Planck for developmental Biology; Alemania
Fil: Reinert, Marina Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
Fil: Vazquez, Martin Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
Materia
Gateway Vector
Nls
P14
Ptrex
Splicing Factor
Trypanosoma
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/79585
Acceder
id CONICETDig_0ffc6f9f51d41b314c4788cf1d184769
oai_identifier_str oai:ri.conicet.gov.ar:11336/79585
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruziWestergaard, Gaston G.Bercovich, NataliaReinert, Marina DanielaVazquez, Martin PabloGateway VectorNlsP14PtrexSplicing FactorTrypanosomahttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1There are only a few reported nuclear localization signals (NLS) in trypanosomes despite intensive research on nuclear metabolic processes such as mRNA processing and transcription during the recent past. Moreover, there are only two reports for a monopartite (La protein) and bipartite (H2B histone, ESAG8) NLS in Trypanosoma brucei. We decided to investigate a NLS in Trypanosoma cruzi by selecting p14, a small RNA recognition motif (RRM) containing protein involved in the splicing process in the nucleus. Its small size (117 amino acids), and an optimized streamlined workflow for analysis in T. cruzi, allowed us to define a region of basic amino acids (RRKRRR) located at the C-terminus that is necessary for nuclear localization. However, the NLS for p14 appeared to be more complex since the signature RRKRRR alone is necessary but not sufficient to direct heterologous proteins, such as GFP, to the nucleus. Since p14 interacts strongly with splicing factor SF3b155, a much larger protein, we designed a p14 variant unable to interact with it. The results allowed us to discard the notion that p14 is entering the nucleus, or is retained within, as the sole consequence of being part of a larger complex. Extensive mapping showed that all of the information for nuclear import resides within the small p14 protein in a bipartite NLS composed of the signature RRKRRR and a region of the RRM domain. Thus, NLS definition in T. cruzi is more complex than previously described.Fil: Westergaard, Gaston G.. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Bercovich, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Max Planck for developmental Biology; AlemaniaFil: Reinert, Marina Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Vazquez, Martin Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaElsevier2010-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/79585Westergaard, Gaston G.; Bercovich, Natalia; Reinert, Marina Daniela; Vazquez, Martin Pablo; Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi; Elsevier; International Journal for Parasitology; 40; 9; 8-2010; 1029-10350020-7519CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S002075191000069Xinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.ijpara.2010.02.011info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:51:15Zoai:ri.conicet.gov.ar:11336/79585instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:51:15.264CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
title Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
spellingShingle Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
Westergaard, Gaston G.
Gateway Vector
Nls
P14
Ptrex
Splicing Factor
Trypanosoma
title_short Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
title_full Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
title_fullStr Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
title_full_unstemmed Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
title_sort Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi
dc.creator.none.fl_str_mv Westergaard, Gaston G.
Bercovich, Natalia
Reinert, Marina Daniela
Vazquez, Martin Pablo
author Westergaard, Gaston G.
author_facet Westergaard, Gaston G.
Bercovich, Natalia
Reinert, Marina Daniela
Vazquez, Martin Pablo
author_role author
author2 Bercovich, Natalia
Reinert, Marina Daniela
Vazquez, Martin Pablo
author2_role author
author
author
dc.subject.none.fl_str_mv Gateway Vector
Nls
P14
Ptrex
Splicing Factor
Trypanosoma
topic Gateway Vector
Nls
P14
Ptrex
Splicing Factor
Trypanosoma
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv There are only a few reported nuclear localization signals (NLS) in trypanosomes despite intensive research on nuclear metabolic processes such as mRNA processing and transcription during the recent past. Moreover, there are only two reports for a monopartite (La protein) and bipartite (H2B histone, ESAG8) NLS in Trypanosoma brucei. We decided to investigate a NLS in Trypanosoma cruzi by selecting p14, a small RNA recognition motif (RRM) containing protein involved in the splicing process in the nucleus. Its small size (117 amino acids), and an optimized streamlined workflow for analysis in T. cruzi, allowed us to define a region of basic amino acids (RRKRRR) located at the C-terminus that is necessary for nuclear localization. However, the NLS for p14 appeared to be more complex since the signature RRKRRR alone is necessary but not sufficient to direct heterologous proteins, such as GFP, to the nucleus. Since p14 interacts strongly with splicing factor SF3b155, a much larger protein, we designed a p14 variant unable to interact with it. The results allowed us to discard the notion that p14 is entering the nucleus, or is retained within, as the sole consequence of being part of a larger complex. Extensive mapping showed that all of the information for nuclear import resides within the small p14 protein in a bipartite NLS composed of the signature RRKRRR and a region of the RRM domain. Thus, NLS definition in T. cruzi is more complex than previously described.
Fil: Westergaard, Gaston G.. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
Fil: Bercovich, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Max Planck for developmental Biology; Alemania
Fil: Reinert, Marina Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
Fil: Vazquez, Martin Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
description There are only a few reported nuclear localization signals (NLS) in trypanosomes despite intensive research on nuclear metabolic processes such as mRNA processing and transcription during the recent past. Moreover, there are only two reports for a monopartite (La protein) and bipartite (H2B histone, ESAG8) NLS in Trypanosoma brucei. We decided to investigate a NLS in Trypanosoma cruzi by selecting p14, a small RNA recognition motif (RRM) containing protein involved in the splicing process in the nucleus. Its small size (117 amino acids), and an optimized streamlined workflow for analysis in T. cruzi, allowed us to define a region of basic amino acids (RRKRRR) located at the C-terminus that is necessary for nuclear localization. However, the NLS for p14 appeared to be more complex since the signature RRKRRR alone is necessary but not sufficient to direct heterologous proteins, such as GFP, to the nucleus. Since p14 interacts strongly with splicing factor SF3b155, a much larger protein, we designed a p14 variant unable to interact with it. The results allowed us to discard the notion that p14 is entering the nucleus, or is retained within, as the sole consequence of being part of a larger complex. Extensive mapping showed that all of the information for nuclear import resides within the small p14 protein in a bipartite NLS composed of the signature RRKRRR and a region of the RRM domain. Thus, NLS definition in T. cruzi is more complex than previously described.
publishDate 2010
dc.date.none.fl_str_mv 2010-08
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/79585
Westergaard, Gaston G.; Bercovich, Natalia; Reinert, Marina Daniela; Vazquez, Martin Pablo; Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi; Elsevier; International Journal for Parasitology; 40; 9; 8-2010; 1029-1035
0020-7519
CONICET Digital
CONICET
url http://hdl.handle.net/11336/79585
identifier_str_mv Westergaard, Gaston G.; Bercovich, Natalia; Reinert, Marina Daniela; Vazquez, Martin Pablo; Analysis of a nuclear localization signal in the p14 splicing factor in Trypanosoma cruzi; Elsevier; International Journal for Parasitology; 40; 9; 8-2010; 1029-1035
0020-7519
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S002075191000069X
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ijpara.2010.02.011
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1846083038074634240
score 13.22299

Publicaciones similares