Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a

Rab GTPases are compartment-specific molecular switches that regulate intracellular vesicular transport in eukaryotes. GDP/GTP exchange factors (GEFs) control Rab activation, and current models propose that localised and regulated GEF activity is important in targeting Rabs to specific membranes. He...

Descripción completa

Detalles Bibliográficos
Autores: Sanzà, Paolo, Evans, Richard D., Briggs, Deborah A., Cantero, Marta, Montoliu, Lluís, Patel, Shyamal, Sviderskaya, Elena V., Itzen, Aymelt, Figueiredo, Ana Cristina, Seabra, Miguel C., Hume, Alistair N.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/240917
Acceso en línea:http://hdl.handle.net/10261/240917
Access Level:acceso abierto
Palabra clave:Organelle transport
Guanine nucleotide exchange factor
Rab27a
Rab3GEP
MADD
Melanocyte
id ES_7d29b2e90bc36802a43c8a68b01f2899
oai_identifier_str oai:digital.csic.es:10261/240917
network_acronym_str ES
network_name_str España
repository_id_str
spelling Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27aSanzà, PaoloEvans, Richard D.Briggs, Deborah A.Cantero, MartaMontoliu, LluísPatel, ShyamalSviderskaya, Elena V.Itzen, AymeltFigueiredo, Ana CristinaSeabra, Miguel C.Hume, Alistair N.Organelle transportGuanine nucleotide exchange factorRab27aRab3GEPMADDMelanocyteRab GTPases are compartment-specific molecular switches that regulate intracellular vesicular transport in eukaryotes. GDP/GTP exchange factors (GEFs) control Rab activation, and current models propose that localised and regulated GEF activity is important in targeting Rabs to specific membranes. Here, we investigated the mechanism of GEF function using the Rab27a GEF, Rab3GEP (also known as MADD), in melanocytes as a model. We show that Rab3GEP-deficient melanocytes (melan-R3GKO) manifest partial disruption of melanosome dispersion, a read-out of Rab27a activation and targeting. Using rescue of melanosome dispersion in melan-R3GKO cells and effector pull-down approaches we show that the DENN domain of Rab3GEP (conserved among RabGEFs) is necessary, but insufficient, for its cellular function and GEF activity. Finally, using a mitochondrial re-targeting strategy, we show that Rab3GEP can target Rab27a to specific membranes in a GEF-dependent manner. We conclude that Rab3GEP facilitates the activation and targeting of Rab27a to specific membranes, but that it differs from other DENN-containing RabGEFs in requiring DENN and non-DENN elements for both of these activities and by lacking compartment-specific localisation.This work was supported by a Medical Research Council New Investigator Award to A.N.H. (grant reference G1100063), Wellcome Trust Project Grant Awards (grant references 091346/Z/10/Z to M.C.S. and A.N.H. and 108429/Z/15/Z to E.V.S.) and a University of Nottingham Vice-Chancellor's PhD Scholarship for Research Excellence (European Union) awarded to P.S. L.M. is funded through the Spanish Ministry of Economy, Industry and Competitiveness (Ministerio de Economía y Competitividad, MINECO) [BIO2015-70978-R]. A.I. is grateful for funding by the Deutsche Forschungsgemeinschaft/Agence Nationale de la Recherche grant IT 91/2-1.Company of BiologistsMedical Research Council (UK)Wellcome TrustUniversity of NottinghamEuropean CommissionMinisterio de Economía y Competitividad (España)German Research FoundationAgence Nationale de la Recherche (France)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2021202120192021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/240917reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2015-70978-Rhttp://dx.doi.org/10.1242/jcs.212035Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2409172026-05-22T06:33:51Z
dc.title.none.fl_str_mv Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
title Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
spellingShingle Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
Sanzà, Paolo
Organelle transport
Guanine nucleotide exchange factor
Rab27a
Rab3GEP
MADD
Melanocyte
title_short Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
title_full Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
title_fullStr Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
title_full_unstemmed Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
title_sort Nucleotide exchange factor Rab3GEP requires DENN and non-DENN elements for activation and targeting of rab27a
dc.creator.none.fl_str_mv Sanzà, Paolo
Evans, Richard D.
Briggs, Deborah A.
Cantero, Marta
Montoliu, Lluís
Patel, Shyamal
Sviderskaya, Elena V.
Itzen, Aymelt
Figueiredo, Ana Cristina
Seabra, Miguel C.
Hume, Alistair N.
author Sanzà, Paolo
author_facet Sanzà, Paolo
Evans, Richard D.
Briggs, Deborah A.
Cantero, Marta
Montoliu, Lluís
Patel, Shyamal
Sviderskaya, Elena V.
Itzen, Aymelt
Figueiredo, Ana Cristina
Seabra, Miguel C.
Hume, Alistair N.
author_role author
author2 Evans, Richard D.
Briggs, Deborah A.
Cantero, Marta
Montoliu, Lluís
Patel, Shyamal
Sviderskaya, Elena V.
Itzen, Aymelt
Figueiredo, Ana Cristina
Seabra, Miguel C.
Hume, Alistair N.
author2_role author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Medical Research Council (UK)
Wellcome Trust
University of Nottingham
European Commission
Ministerio de Economía y Competitividad (España)
German Research Foundation
Agence Nationale de la Recherche (France)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Organelle transport
Guanine nucleotide exchange factor
Rab27a
Rab3GEP
MADD
Melanocyte
topic Organelle transport
Guanine nucleotide exchange factor
Rab27a
Rab3GEP
MADD
Melanocyte
description Rab GTPases are compartment-specific molecular switches that regulate intracellular vesicular transport in eukaryotes. GDP/GTP exchange factors (GEFs) control Rab activation, and current models propose that localised and regulated GEF activity is important in targeting Rabs to specific membranes. Here, we investigated the mechanism of GEF function using the Rab27a GEF, Rab3GEP (also known as MADD), in melanocytes as a model. We show that Rab3GEP-deficient melanocytes (melan-R3GKO) manifest partial disruption of melanosome dispersion, a read-out of Rab27a activation and targeting. Using rescue of melanosome dispersion in melan-R3GKO cells and effector pull-down approaches we show that the DENN domain of Rab3GEP (conserved among RabGEFs) is necessary, but insufficient, for its cellular function and GEF activity. Finally, using a mitochondrial re-targeting strategy, we show that Rab3GEP can target Rab27a to specific membranes in a GEF-dependent manner. We conclude that Rab3GEP facilitates the activation and targeting of Rab27a to specific membranes, but that it differs from other DENN-containing RabGEFs in requiring DENN and non-DENN elements for both of these activities and by lacking compartment-specific localisation.
publishDate 2019
dc.date.none.fl_str_mv 2019
2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/240917
url http://hdl.handle.net/10261/240917
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2015-70978-R
http://dx.doi.org/10.1242/jcs.212035
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Company of Biologists
publisher.none.fl_str_mv Company of Biologists
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869411643417427968
score 15,81155

Similares en LA Referencia