SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing

Pre-mRNA splicing is catalyzed by the spliceosome, a multi-megadalton ribonucleoprotein machine. Previous work from our laboratory revealed the splicing factor SRSF1 as a regulator of the SUMO pathway, leading us to explore a connection between this pathway and the splicing machinery. We show here t...

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Detalhes bibliográficos
Autores: Pozzi, María Berta, Bragado, Laureano Fabian Tomas, Will, Cindy L., Mammi, Pablo Andrés, Risso, Guillermo, Urlaub, Henning, Lührmann, Reinhard, Srebrow, Anabella
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2017
País:Argentina
Recursos:Consejo Nacional de Investigaciones Científicas y Técnicas
Repositorio:CONICET Digital (CONICET)
Idioma:inglés
OAI Identifier:oai:ri.conicet.gov.ar:11336/65300
Acesso em linha:http://hdl.handle.net/11336/65300
Access Level:acceso abierto
Palavra-chave:SUMO
SPLICING
PRP3
SPLICEOSOME
SRSF1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
id AR_913d9d65274829c59bfdbf7d5ea0fc66
oai_identifier_str oai:ri.conicet.gov.ar:11336/65300
network_acronym_str AR
network_name_str Argentina
repository_id_str
dc.title.none.fl_str_mv SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
title SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
spellingShingle SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
Pozzi, María Berta
SUMO
SPLICING
PRP3
SPLICEOSOME
SRSF1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
title_short SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
title_full SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
title_fullStr SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
title_full_unstemmed SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
title_sort SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing
dc.creator.none.fl_str_mv Pozzi, María Berta
Bragado, Laureano Fabian Tomas
Will, Cindy L.
Mammi, Pablo Andrés
Risso, Guillermo
Urlaub, Henning
Lührmann, Reinhard
Srebrow, Anabella
author Pozzi, María Berta
author_facet Pozzi, María Berta
Bragado, Laureano Fabian Tomas
Will, Cindy L.
Mammi, Pablo Andrés
Risso, Guillermo
Urlaub, Henning
Lührmann, Reinhard
Srebrow, Anabella
author_role author
author2 Bragado, Laureano Fabian Tomas
Will, Cindy L.
Mammi, Pablo Andrés
Risso, Guillermo
Urlaub, Henning
Lührmann, Reinhard
Srebrow, Anabella
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv SUMO
SPLICING
PRP3
SPLICEOSOME
SRSF1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
topic SUMO
SPLICING
PRP3
SPLICEOSOME
SRSF1
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
description Pre-mRNA splicing is catalyzed by the spliceosome, a multi-megadalton ribonucleoprotein machine. Previous work from our laboratory revealed the splicing factor SRSF1 as a regulator of the SUMO pathway, leading us to explore a connection between this pathway and the splicing machinery. We show here that addition of a recombinant SUMO-protease decreases the efficiency of pre-mRNA splicing in vitro. By mass spectrometry analysis of anti-SUMO immunoprecipitated proteins obtained from purified splicing complexes formed along the splicing reaction, we identified spliceosome-associated SUMO substrates. After corroborating SUMOylation of Prp3 in cultured cells, we defined Lys 289 and Lys 559 as bona fide SUMO attachment sites within this spliceosomal protein. We further demonstrated that a Prp3 SUMOylation-deficient mutant while still capable of interacting with U4/U6 snRNP components, is unable to co-precipitate U2 and U5 snRNA and the spliceosomal proteins U2-SF3a120 and U5-Snu114. This SUMOylation-deficient mutant fails to restore the splicing of different pre-mRNAs to the levels achieved by the wild type protein, when transfected into Prp3-depleted cultured cells. This mutant also shows a diminished recruitment to active spliceosomes, compared to the wild type protein. These findings indicate that SUMO conjugation plays a role during the splicing process and suggest the involvement of Prp3 SUMOylation in U4/U6U5 tri-snRNP formation and/or recruitment.
publishDate 2017
dc.date.none.fl_str_mv 2017-06
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/11336/65300
Pozzi, María Berta; Bragado, Laureano Fabian Tomas; Will, Cindy L.; Mammi, Pablo Andrés; Risso, Guillermo; et al.; SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing; Oxford University Press; Nucleic Acids Research; 45; 11; 6-2017; 6729-6745
0305-1048
1362-4962
CONICET Digital
CONICET
url http://hdl.handle.net/11336/65300
identifier_str_mv Pozzi, María Berta; Bragado, Laureano Fabian Tomas; Will, Cindy L.; Mammi, Pablo Andrés; Risso, Guillermo; et al.; SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing; Oxford University Press; Nucleic Acids Research; 45; 11; 6-2017; 6729-6745
0305-1048
1362-4962
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1093/nar/gkx213
info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/nar/article/45/11/6729/3096161
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
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
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_ 1799195947708710912
spelling SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicingPozzi, María BertaBragado, Laureano Fabian TomasWill, Cindy L.Mammi, Pablo AndrésRisso, GuillermoUrlaub, HenningLührmann, ReinhardSrebrow, AnabellaSUMOSPLICINGPRP3SPLICEOSOMESRSF1https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Pre-mRNA splicing is catalyzed by the spliceosome, a multi-megadalton ribonucleoprotein machine. Previous work from our laboratory revealed the splicing factor SRSF1 as a regulator of the SUMO pathway, leading us to explore a connection between this pathway and the splicing machinery. We show here that addition of a recombinant SUMO-protease decreases the efficiency of pre-mRNA splicing in vitro. By mass spectrometry analysis of anti-SUMO immunoprecipitated proteins obtained from purified splicing complexes formed along the splicing reaction, we identified spliceosome-associated SUMO substrates. After corroborating SUMOylation of Prp3 in cultured cells, we defined Lys 289 and Lys 559 as bona fide SUMO attachment sites within this spliceosomal protein. We further demonstrated that a Prp3 SUMOylation-deficient mutant while still capable of interacting with U4/U6 snRNP components, is unable to co-precipitate U2 and U5 snRNA and the spliceosomal proteins U2-SF3a120 and U5-Snu114. This SUMOylation-deficient mutant fails to restore the splicing of different pre-mRNAs to the levels achieved by the wild type protein, when transfected into Prp3-depleted cultured cells. This mutant also shows a diminished recruitment to active spliceosomes, compared to the wild type protein. These findings indicate that SUMO conjugation plays a role during the splicing process and suggest the involvement of Prp3 SUMOylation in U4/U6U5 tri-snRNP formation and/or recruitment.Fil: Pozzi, María Berta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Bragado, Laureano Fabian Tomas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Will, Cindy L.. Max Planck Institute For Biophysical Chemistry; AlemaniaFil: Mammi, Pablo Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Risso, Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Urlaub, Henning. Max Planck Institute For Biophysical Chemistry; AlemaniaFil: Lührmann, Reinhard. Max Planck Institute For Biophysical Chemistry; AlemaniaFil: Srebrow, Anabella. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaOxford University Press2017-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/65300Pozzi, María Berta; Bragado, Laureano Fabian Tomas; Will, Cindy L.; Mammi, Pablo Andrés; Risso, Guillermo; et al.; SUMO conjugation to spliceosomal proteins is required for efficient pre-mRNA splicing; Oxford University Press; Nucleic Acids Research; 45; 11; 6-2017; 6729-67450305-10481362-4962CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1093/nar/gkx213info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/nar/article/45/11/6729/3096161info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2024-05-08T14:09:10Zoai:ri.conicet.gov.ar:11336/65300instacron: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:34982024-05-08 14:09:11.204CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
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