Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain

PMCID: PMC5609853

Detalhes bibliográficos
Autores: Boettner, Douglas R., Segarra, Verónica A., Moorthy, Balaji T., León, Nagore de, Creagh, John, Collette, John R., Malhotra, Arun, Lemmon, Sandra K.
Formato: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2016
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/156914
Acesso em linha:http://hdl.handle.net/10261/156914
Access Level:acceso abierto
Palavra-chave:TGN/endosomal sorting
Endocytosis
Clathrin
Membrane trafficking
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spelling Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chainBoettner, Douglas R.Segarra, Verónica A.Moorthy, Balaji T.León, Nagore deCreagh, JohnCollette, John R.Malhotra, ArunLemmon, Sandra K.TGN/endosomal sortingEndocytosisClathrinMembrane traffickingPMCID: PMC5609853Clathrin facilitates vesicle formation during endocytosis and sorting in the trans-Golgi network (TGN)/endosomal system. Unlike in mammals, yeast clathrin function requires both the clathrin heavy (CHC) and clathrin light (CLC) chain, since Chc1 does not form stable trimers without Clc1. To further delineate clathrin subunit functions, we constructed a chimeric CHC protein (Chc-YR), which fused the N-terminus of yeast CHC (1–1312) to the rat CHC residues 1318–1675, including the CHC trimerization region. The novel CHC-YR allele encoded a stable protein that fractionated as a trimer. CHC-YR also complemented chc1Δ slow growth and clathrin TGN/endosomal sorting defects. In strains depleted for Clc1 (either clc1Δ or chc1Δ clc1Δ), CHC-YR, but not CHC1, suppressed TGN/endosomal sorting and growth phenotypes. Chc-YR-GFP (green fluorescent protein) localized to the TGN and cortical patches on the plasma membrane, like Chc1 and Clc1. However, Clc1-GFP was primarily cytoplasmic in chc1Δ cells harboring pCHC-YR, indicating that Chc-YR does not bind yeast CLC. Still, some partial phenotypes persisted in cells with Chc-YR, which are likely due either to loss of CLC recruitment or chimeric HC lattice instability. Ultimately, these studies have created a tool to examine non-trimerization roles for the clathrin LC.This work was supported primarily by the National Institute of Health grants to SKL (R01-GM055796), DRB (F32-GM084677) and an institutional training grant supporting VAS (T32-HL07188). Further support was provided by the American Cancer Society fellowship to JC (FL Summer Research Fellowship), American Heart Association Postdoctoral fellowship to JRC (6-6253Y), and a European Molecular Biology Organization Short Term Fellowship to ND (EMBO145-2012).Peer ReviewedJohn Wiley & SonsEMBOAmerican Heart AssociationAmerican Cancer SocietyNational Institutes of Health (US)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2017201720162017info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionhttp://hdl.handle.net/10261/156914reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://doi.org/10.1111/tra.12401Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1569142026-05-22T06:33:51Z
dc.title.none.fl_str_mv Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
title Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
spellingShingle Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
Boettner, Douglas R.
TGN/endosomal sorting
Endocytosis
Clathrin
Membrane trafficking
title_short Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
title_full Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
title_fullStr Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
title_full_unstemmed Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
title_sort Creating a chimeric clathrin heavy chain that functions independently of yeast clathrin light chain
dc.creator.none.fl_str_mv Boettner, Douglas R.
Segarra, Verónica A.
Moorthy, Balaji T.
León, Nagore de
Creagh, John
Collette, John R.
Malhotra, Arun
Lemmon, Sandra K.
author Boettner, Douglas R.
author_facet Boettner, Douglas R.
Segarra, Verónica A.
Moorthy, Balaji T.
León, Nagore de
Creagh, John
Collette, John R.
Malhotra, Arun
Lemmon, Sandra K.
author_role author
author2 Segarra, Verónica A.
Moorthy, Balaji T.
León, Nagore de
Creagh, John
Collette, John R.
Malhotra, Arun
Lemmon, Sandra K.
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv EMBO
American Heart Association
American Cancer Society
National Institutes of Health (US)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv TGN/endosomal sorting
Endocytosis
Clathrin
Membrane trafficking
topic TGN/endosomal sorting
Endocytosis
Clathrin
Membrane trafficking
description PMCID: PMC5609853
publishDate 2016
dc.date.none.fl_str_mv 2016
2017
2017
2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Postprint
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/156914
url http://hdl.handle.net/10261/156914
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://doi.org/10.1111/tra.12401
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv John Wiley & Sons
publisher.none.fl_str_mv John Wiley & Sons
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_ 1869408181885599744
score 15.812429

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