Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1

Autores
Vazquez, Martin Pablo; Mualem, David; Bercovich, Natalia; Stern, Michael Zeev; Nyambega, Benson; Barda, Omer; Masiga, Dan; Gupta, Sachin Kumar; Michaeli, Shulamit; Levin, Mariano Jorge
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, we identified, cloned and expressed the Trypanosoma cruzi and Trypanosoma brucei U2AF35, U2AF65 and SF1. Trypanosomatid U2AF65 strongly diverged from yeast and human homologues. On the contrary, trypanosomatid SF1 was conserved but lacked the C-terminal sequence present in the mammalian protein. Yeast two hybrid approaches were used to assess their interactions. The interaction between U2AF35 and U2AF65 was very weak or not detectable. However, as in other eukaryotes, the interaction between U2AF65 and SF1 was strong. At the cellular level, these results were confirmed by fractionation and affinity-selection experiments in which SF1 and U2AF65 co-fractionated in a complex of approximately 400 kDa and U2AF65 was affinity-selected with TAP tagged SF1, but not with TAP tagged U2AF35. Silencing of the three factors affected growth and trans-splicing in the first step of this reaction. Trypanosomes are the first described example of eukaryotic cells in which the interaction of two expressed U2AF factors seemed to be very weak, or even undetectable.
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
Fil: Mualem, David. Bar-Ilan University; Israel
Fil: Bercovich, Natalia. 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
Fil: Stern, Michael Zeev. Bar-Ilan University; Israel
Fil: Nyambega, Benson. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. International Center for Insect Physiology and Ecology; Kenia
Fil: Barda, Omer. Bar-Ilan University; Israel
Fil: Masiga, Dan. International Center for Insect Physiology and Ecology; Kenia
Fil: Gupta, Sachin Kumar. Bar-Ilan University; Israel
Fil: Michaeli, Shulamit. Bar-Ilan University; Israel
Fil: Levin, Mariano Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Inserm; Francia
Materia
Trans-Splicing
Trypanosoma Brucei
Trypanosoma Cruzi
U2 Auxiliary Factor
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:11336/79490
Acceder
id CONICETDig_123b2e1c4d7782de3989d63adb8346f7
oai_identifier_str oai:ri.conicet.gov.ar:11336/79490
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1Vazquez, Martin PabloMualem, DavidBercovich, NataliaStern, Michael ZeevNyambega, BensonBarda, OmerMasiga, DanGupta, Sachin KumarMichaeli, ShulamitLevin, Mariano JorgeTrans-SplicingTrypanosoma BruceiTrypanosoma CruziU2 Auxiliary Factorhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, we identified, cloned and expressed the Trypanosoma cruzi and Trypanosoma brucei U2AF35, U2AF65 and SF1. Trypanosomatid U2AF65 strongly diverged from yeast and human homologues. On the contrary, trypanosomatid SF1 was conserved but lacked the C-terminal sequence present in the mammalian protein. Yeast two hybrid approaches were used to assess their interactions. The interaction between U2AF35 and U2AF65 was very weak or not detectable. However, as in other eukaryotes, the interaction between U2AF65 and SF1 was strong. At the cellular level, these results were confirmed by fractionation and affinity-selection experiments in which SF1 and U2AF65 co-fractionated in a complex of approximately 400 kDa and U2AF65 was affinity-selected with TAP tagged SF1, but not with TAP tagged U2AF35. Silencing of the three factors affected growth and trans-splicing in the first step of this reaction. Trypanosomes are the first described example of eukaryotic cells in which the interaction of two expressed U2AF factors seemed to be very weak, or even undetectable.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; ArgentinaFil: Mualem, David. Bar-Ilan University; IsraelFil: Bercovich, Natalia. 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; ArgentinaFil: Stern, Michael Zeev. Bar-Ilan University; IsraelFil: Nyambega, Benson. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. International Center for Insect Physiology and Ecology; KeniaFil: Barda, Omer. Bar-Ilan University; IsraelFil: Masiga, Dan. International Center for Insect Physiology and Ecology; KeniaFil: Gupta, Sachin Kumar. Bar-Ilan University; IsraelFil: Michaeli, Shulamit. Bar-Ilan University; IsraelFil: Levin, Mariano Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Inserm; FranciaElsevier Science2009-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/79490Vazquez, Martin Pablo; Mualem, David; Bercovich, Natalia; Stern, Michael Zeev; Nyambega, Benson; et al.; Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1; Elsevier Science; Molecular and Biochemical Parasitology; 164; 2; 4-2009; 137-1460166-6851CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.molbiopara.2008.12.009info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0166685109000024info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:31:12Zoai:ri.conicet.gov.ar:11336/79490instacron: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:34982025-09-29 10:31:13.125CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
title Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
spellingShingle Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
Vazquez, Martin Pablo
Trans-Splicing
Trypanosoma Brucei
Trypanosoma Cruzi
U2 Auxiliary Factor
title_short Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
title_full Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
title_fullStr Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
title_full_unstemmed Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
title_sort Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1
dc.creator.none.fl_str_mv Vazquez, Martin Pablo
Mualem, David
Bercovich, Natalia
Stern, Michael Zeev
Nyambega, Benson
Barda, Omer
Masiga, Dan
Gupta, Sachin Kumar
Michaeli, Shulamit
Levin, Mariano Jorge
author Vazquez, Martin Pablo
author_facet Vazquez, Martin Pablo
Mualem, David
Bercovich, Natalia
Stern, Michael Zeev
Nyambega, Benson
Barda, Omer
Masiga, Dan
Gupta, Sachin Kumar
Michaeli, Shulamit
Levin, Mariano Jorge
author_role author
author2 Mualem, David
Bercovich, Natalia
Stern, Michael Zeev
Nyambega, Benson
Barda, Omer
Masiga, Dan
Gupta, Sachin Kumar
Michaeli, Shulamit
Levin, Mariano Jorge
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Trans-Splicing
Trypanosoma Brucei
Trypanosoma Cruzi
U2 Auxiliary Factor
topic Trans-Splicing
Trypanosoma Brucei
Trypanosoma Cruzi
U2 Auxiliary Factor
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, we identified, cloned and expressed the Trypanosoma cruzi and Trypanosoma brucei U2AF35, U2AF65 and SF1. Trypanosomatid U2AF65 strongly diverged from yeast and human homologues. On the contrary, trypanosomatid SF1 was conserved but lacked the C-terminal sequence present in the mammalian protein. Yeast two hybrid approaches were used to assess their interactions. The interaction between U2AF35 and U2AF65 was very weak or not detectable. However, as in other eukaryotes, the interaction between U2AF65 and SF1 was strong. At the cellular level, these results were confirmed by fractionation and affinity-selection experiments in which SF1 and U2AF65 co-fractionated in a complex of approximately 400 kDa and U2AF65 was affinity-selected with TAP tagged SF1, but not with TAP tagged U2AF35. Silencing of the three factors affected growth and trans-splicing in the first step of this reaction. Trypanosomes are the first described example of eukaryotic cells in which the interaction of two expressed U2AF factors seemed to be very weak, or even undetectable.
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
Fil: Mualem, David. Bar-Ilan University; Israel
Fil: Bercovich, Natalia. 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
Fil: Stern, Michael Zeev. Bar-Ilan University; Israel
Fil: Nyambega, Benson. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. International Center for Insect Physiology and Ecology; Kenia
Fil: Barda, Omer. Bar-Ilan University; Israel
Fil: Masiga, Dan. International Center for Insect Physiology and Ecology; Kenia
Fil: Gupta, Sachin Kumar. Bar-Ilan University; Israel
Fil: Michaeli, Shulamit. Bar-Ilan University; Israel
Fil: Levin, Mariano Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Inserm; Francia
description Early in the assembly of the spliceosome of eukaryotes the branch-point binding protein (BBP, also called SF1) recognizes the branch point sequence, whereas the heterodimer U2AF consisting of a 65 and a 35 kDa subunit, contacts the polypyrimidine tract and the AG splice site, respectively. Herein, we identified, cloned and expressed the Trypanosoma cruzi and Trypanosoma brucei U2AF35, U2AF65 and SF1. Trypanosomatid U2AF65 strongly diverged from yeast and human homologues. On the contrary, trypanosomatid SF1 was conserved but lacked the C-terminal sequence present in the mammalian protein. Yeast two hybrid approaches were used to assess their interactions. The interaction between U2AF35 and U2AF65 was very weak or not detectable. However, as in other eukaryotes, the interaction between U2AF65 and SF1 was strong. At the cellular level, these results were confirmed by fractionation and affinity-selection experiments in which SF1 and U2AF65 co-fractionated in a complex of approximately 400 kDa and U2AF65 was affinity-selected with TAP tagged SF1, but not with TAP tagged U2AF35. Silencing of the three factors affected growth and trans-splicing in the first step of this reaction. Trypanosomes are the first described example of eukaryotic cells in which the interaction of two expressed U2AF factors seemed to be very weak, or even undetectable.
publishDate 2009
dc.date.none.fl_str_mv 2009-04
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/11336/79490
Vazquez, Martin Pablo; Mualem, David; Bercovich, Natalia; Stern, Michael Zeev; Nyambega, Benson; et al.; Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1; Elsevier Science; Molecular and Biochemical Parasitology; 164; 2; 4-2009; 137-146
0166-6851
CONICET Digital
CONICET
url http://hdl.handle.net/11336/79490
identifier_str_mv Vazquez, Martin Pablo; Mualem, David; Bercovich, Natalia; Stern, Michael Zeev; Nyambega, Benson; et al.; Functional characterization and protein-protein interactions of trypanosome splicing factors U2AF35, U2AF65 and SF1; Elsevier Science; Molecular and Biochemical Parasitology; 164; 2; 4-2009; 137-146
0166-6851
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.molbiopara.2008.12.009
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0166685109000024
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
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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_ 1844614322149392384
score 13.070432

Publicaciones similares