WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1

Detalhes bibliográficos
Autor(a) principal: Henriques, Andreia F.A.
Data de Publicação: 2020
Outros Autores: Matos, Paulo, AS, Carvalho, Azkargorta, Mikel, Elortza, Felix, Matthiesen, Rune, Jordan, Peter
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/155151
Resumo: This work was supported by Fundação para a Ciência e Tecnologia (FCT) [grants PTDC/SAU-MET/117236/2010 to PJ, grant UID/MULTI/ 04046/2019 to the research unit BioISI, and fellowship SFRH/BD/ 106080/2015 from the BioSYS PhD programme PD65-2012 to AFAH]. The authors acknowledge José Ferrão for regular mycoplasma testing in cultured cells and Patrícia Barros for critical reading of the manuscript. Anthony Albiston (Melbourne, Australia) and Florian Lang (University of Tubingen, Germany) kindly provided constucts pCR3.1/AS160-2myc or pIRES2-EGFP-SGK1, respectively, for this study.
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spelling WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1Glucose cotransportGLUT1Membrane trafficProtein phosphorylationTBC1DWNK1BiophysicsBiochemistryMolecular BiologySDG 3 - Good Health and Well-beingThis work was supported by Fundação para a Ciência e Tecnologia (FCT) [grants PTDC/SAU-MET/117236/2010 to PJ, grant UID/MULTI/ 04046/2019 to the research unit BioISI, and fellowship SFRH/BD/ 106080/2015 from the BioSYS PhD programme PD65-2012 to AFAH]. The authors acknowledge José Ferrão for regular mycoplasma testing in cultured cells and Patrícia Barros for critical reading of the manuscript. Anthony Albiston (Melbourne, Australia) and Florian Lang (University of Tubingen, Germany) kindly provided constucts pCR3.1/AS160-2myc or pIRES2-EGFP-SGK1, respectively, for this study.Glucose uptake by mammalian cells is a key mechanism to maintain cell and tissue homeostasis and relies mostly on plasma membrane-localized glucose transporter proteins (GLUTs). Two main cellular mechanisms regulate GLUT proteins in the cell: first, expression of GLUT genes is under dynamic transcriptional control and is used by cancer cells to increase glucose availability. Second, GLUT proteins are regulated by membrane traffic from storage vesicles to the plasma membrane (PM). This latter process is triggered by signaling mechanisms and well-studied in the case of insulin-responsive cells, which activate protein kinase AKT to phosphorylate TBC1D4, a RAB-GTPase activating protein involved in membrane traffic regulation. Previously, we identified protein kinase WNK1 as another kinase able to phosphorylate TBC1D4 and regulate the surface expression of the constitutive glucose transporter GLUT1. Here we describe that downregulation of WNK1 through RNA interference in HEK293 cells led to a 2-fold decrease in PM GLUT1 expression, concomitant with a 60% decrease in glucose uptake. By mass spectrometry, we identified serine (S) 704 in TBC1D4 as a WNK1-regulated phosphorylation site, and also S565 in the paralogue TBC1D1. Transfection of the respective phosphomimetic or unphosphorylatable TBC1D mutants into cells revealed that both affected the cell surface abundance of GLUT1. The results reinforce a regulatory role for WNK1 in cell metabolism and have potential impact for the understanding of cancer cell metabolism and therapeutic options in type 2 diabetes.NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)Centro de Estudos de Doenças Crónicas (CEDOC)RUNHenriques, Andreia F.A.Matos, PauloAS, CarvalhoAzkargorta, MikelElortza, FelixMatthiesen, RuneJordan, Peter2023-07-12T22:13:24Z2020-01-152020-01-15T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/155151eng0003-9861PURE: 15926503https://doi.org/10.1016/j.abb.2019.108223info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:37:38Zoai:run.unl.pt:10362/155151Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:55:55.951330Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
title WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
spellingShingle WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
Henriques, Andreia F.A.
Glucose cotransport
GLUT1
Membrane traffic
Protein phosphorylation
TBC1D
WNK1
Biophysics
Biochemistry
Molecular Biology
SDG 3 - Good Health and Well-being
title_short WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
title_full WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
title_fullStr WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
title_full_unstemmed WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
title_sort WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
author Henriques, Andreia F.A.
author_facet Henriques, Andreia F.A.
Matos, Paulo
AS, Carvalho
Azkargorta, Mikel
Elortza, Felix
Matthiesen, Rune
Jordan, Peter
author_role author
author2 Matos, Paulo
AS, Carvalho
Azkargorta, Mikel
Elortza, Felix
Matthiesen, Rune
Jordan, Peter
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM)
Centro de Estudos de Doenças Crónicas (CEDOC)
RUN
dc.contributor.author.fl_str_mv Henriques, Andreia F.A.
Matos, Paulo
AS, Carvalho
Azkargorta, Mikel
Elortza, Felix
Matthiesen, Rune
Jordan, Peter
dc.subject.por.fl_str_mv Glucose cotransport
GLUT1
Membrane traffic
Protein phosphorylation
TBC1D
WNK1
Biophysics
Biochemistry
Molecular Biology
SDG 3 - Good Health and Well-being
topic Glucose cotransport
GLUT1
Membrane traffic
Protein phosphorylation
TBC1D
WNK1
Biophysics
Biochemistry
Molecular Biology
SDG 3 - Good Health and Well-being
description This work was supported by Fundação para a Ciência e Tecnologia (FCT) [grants PTDC/SAU-MET/117236/2010 to PJ, grant UID/MULTI/ 04046/2019 to the research unit BioISI, and fellowship SFRH/BD/ 106080/2015 from the BioSYS PhD programme PD65-2012 to AFAH]. The authors acknowledge José Ferrão for regular mycoplasma testing in cultured cells and Patrícia Barros for critical reading of the manuscript. Anthony Albiston (Melbourne, Australia) and Florian Lang (University of Tubingen, Germany) kindly provided constucts pCR3.1/AS160-2myc or pIRES2-EGFP-SGK1, respectively, for this study.
publishDate 2020
dc.date.none.fl_str_mv 2020-01-15
2020-01-15T00:00:00Z
2023-07-12T22:13:24Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/155151
url http://hdl.handle.net/10362/155151
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0003-9861
PURE: 15926503
https://doi.org/10.1016/j.abb.2019.108223
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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