The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein

Sos1 is an universal, widely expressed Ras guanine nucleotide-exchange factor (RasGEF) in eukaryotic cells. Its N-terminal HD motif is known to be involved in allosteric regulation of Sos1 GEF activity through intramolecular interaction with the neighboring PH domain. Here, we searched for other cel...

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Autores: Zarich-Dimitrievich, Natasha, Anta, Begoña, Fernández-Medarde, Alberto, Ballester, Alicia, de Lucas, Maria Pilar, Camara, Ana Belen, Anta-Felez, Berta, Oliva-Martinez, Jose Luis, Rojas-Cabañeros, Jose Maria, Santos, Eugenio
Tipo de recurso: artículo
Fecha de publicación:2019
País:España
Institución:Instituto de Salud Carlos III (ISCIII)
Repositorio:Repisalud
Idioma:inglés
OAI Identifier:oai:repisalud.isciii.es:20.500.12105/7441
Acceso en línea:http://hdl.handle.net/20.500.12105/7441
Access Level:acceso abierto
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spelling The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF proteinZarich-Dimitrievich, NatashaAnta, BegoñaFernández-Medarde, AlbertoBallester, Aliciade Lucas, Maria PilarCamara, Ana BelenAnta-Felez, BertaOliva-Martinez, Jose LuisRojas-Cabañeros, Jose MariaSantos, EugenioSos1 is an universal, widely expressed Ras guanine nucleotide-exchange factor (RasGEF) in eukaryotic cells. Its N-terminal HD motif is known to be involved in allosteric regulation of Sos1 GEF activity through intramolecular interaction with the neighboring PH domain. Here, we searched for other cellular proteins also able to interact productively with the Sos1 HD domain. Using a yeast two-hybrid system, we identified the interaction between the Sos1 HD region and CSN3, the third component of the COP9 signalosome, a conserved, multi-subunit protein complex that functions in the ubiquitin-proteasome pathway to control degradation of many cellular proteins. The interaction of CSN3 with the HD of Sos1 was confirmed in vitro by GST pull-down assays using truncated mutants and reproduced in vivo by co-immunoprecipitation with the endogenous, full-length cellular Sos1 protein. In vitro kinase assays showed that PKD, a COP9 signalosome-associated-kinase, is able to phosphorylate Sos1. The intracellular levels of Sos1 protein were clearly diminished following CSN3 or PKD knockdown. A sizable fraction of the endogenous Sos1 protein was found ubiquitinated in different mammalian cell types. A significant reduction of RasGTP formation upon growth factor stimulation was also observed in CSN3-silenced as compared with control cells. Our data suggest that the interaction of Sos1 with the COP9 signalosome and PKD plays a significant role in maintenance of cellular Sos1 protein stability and homeostasis under physiological conditions and raises the possibility of considering the CSN/PKD complex as a potential target for design of novel therapeutic drugs.Nature Publishing GroupJunta de Castilla y León (España)Fundación Memoria de Don Samuel SolórzanoInstituto de Salud Carlos IIIAsociación Española Contra el CáncerMinisterio de Economía y Competitividad (España)Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)20192019-04-0920192019-01-0420192019-01-04research articlehttp://purl.org/coar/resource_type/c_2df8fbb1VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/20.500.12105/7441reponame:Repisaludinstname:Instituto de Salud Carlos III (ISCIII)InglésengES SAF2016-78852-R Not availableES PI13 00703ES PI16 02137ES RD12 0036ES RD12 0036open accesshttp://purl.org/coar/access_right/c_abf2Atribución 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repisalud.isciii.es:20.500.12105/74412026-06-12T12:43:37Z
dc.title.none.fl_str_mv The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
title The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
spellingShingle The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
Zarich-Dimitrievich, Natasha
title_short The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
title_full The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
title_fullStr The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
title_full_unstemmed The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
title_sort The CSN3 subunit of the COP9 signalosome interacts with the HD region of Sos1 regulating stability of this GEF protein
dc.creator.none.fl_str_mv Zarich-Dimitrievich, Natasha
Anta, Begoña
Fernández-Medarde, Alberto
Ballester, Alicia
de Lucas, Maria Pilar
Camara, Ana Belen
Anta-Felez, Berta
Oliva-Martinez, Jose Luis
Rojas-Cabañeros, Jose Maria
Santos, Eugenio
author Zarich-Dimitrievich, Natasha
author_facet Zarich-Dimitrievich, Natasha
Anta, Begoña
Fernández-Medarde, Alberto
Ballester, Alicia
de Lucas, Maria Pilar
Camara, Ana Belen
Anta-Felez, Berta
Oliva-Martinez, Jose Luis
Rojas-Cabañeros, Jose Maria
Santos, Eugenio
author_role author
author2 Anta, Begoña
Fernández-Medarde, Alberto
Ballester, Alicia
de Lucas, Maria Pilar
Camara, Ana Belen
Anta-Felez, Berta
Oliva-Martinez, Jose Luis
Rojas-Cabañeros, Jose Maria
Santos, Eugenio
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Junta de Castilla y León (España)
Fundación Memoria de Don Samuel Solórzano
Instituto de Salud Carlos III
Asociación Española Contra el Cáncer
Ministerio de Economía y Competitividad (España)
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
description Sos1 is an universal, widely expressed Ras guanine nucleotide-exchange factor (RasGEF) in eukaryotic cells. Its N-terminal HD motif is known to be involved in allosteric regulation of Sos1 GEF activity through intramolecular interaction with the neighboring PH domain. Here, we searched for other cellular proteins also able to interact productively with the Sos1 HD domain. Using a yeast two-hybrid system, we identified the interaction between the Sos1 HD region and CSN3, the third component of the COP9 signalosome, a conserved, multi-subunit protein complex that functions in the ubiquitin-proteasome pathway to control degradation of many cellular proteins. The interaction of CSN3 with the HD of Sos1 was confirmed in vitro by GST pull-down assays using truncated mutants and reproduced in vivo by co-immunoprecipitation with the endogenous, full-length cellular Sos1 protein. In vitro kinase assays showed that PKD, a COP9 signalosome-associated-kinase, is able to phosphorylate Sos1. The intracellular levels of Sos1 protein were clearly diminished following CSN3 or PKD knockdown. A sizable fraction of the endogenous Sos1 protein was found ubiquitinated in different mammalian cell types. A significant reduction of RasGTP formation upon growth factor stimulation was also observed in CSN3-silenced as compared with control cells. Our data suggest that the interaction of Sos1 with the COP9 signalosome and PKD plays a significant role in maintenance of cellular Sos1 protein stability and homeostasis under physiological conditions and raises the possibility of considering the CSN/PKD complex as a potential target for design of novel therapeutic drugs.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019-04-09
2019
2019-01-04
2019
2019-01-04
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12105/7441
url http://hdl.handle.net/20.500.12105/7441
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv ES SAF2016-78852-R Not available
ES PI13 00703
ES PI16 02137
ES RD12 0036
ES RD12 0036
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 4.0 Internacional
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 4.0 Internacional
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:Repisalud
instname:Instituto de Salud Carlos III (ISCIII)
instname_str Instituto de Salud Carlos III (ISCIII)
reponame_str Repisalud
collection Repisalud
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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