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Metabolic tumor cell adaptation: tyrosine phosphorylation modulates cell surface expression of NKCC2 and KCC3

2020-03-03Conference object Open access
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dc.contributor.authorLoureiro, Cláudia
dc.contributor.authorBarros, Patrícia
dc.contributor.authorMatos, Paulo
dc.contributor.authorJordan, Peter
dc.date.accessioned2021-03-13T09:25:12Z
dc.date.available2021-03-13T09:25:12Z
dc.date.issued2020-03-03
dc.description.abstractIntroduction: Tumor cells require cellular chloride and potassium transport to adapt to a changing microenvironment, both for cell volume regulation and membrane potential maintenance. Cellular chloride and potassium entry or exit are mediated at the plasma membrane by cotransporter proteins of the solute carrier 12 family. For example, NKCC2 resorbs chloride with sodium and potassium ions at the apical membrane of epithelial cells in the kidney, whereas KCC3 releases chloride with potassium ions at the basolateral membrane. Their ion transport activity is regulated by protein phosphorylation in response to signaling pathways. An additional regulatory mechanism concerns the amount of cotransporter molecules inserted into the plasma membrane. Material and Methods: Cotransporter constructs were transfected into HEK293 cells and the activity of spleen tyrosine kinase (SYK) modulated by incubation with SYK inhibitors or by co-transfection with siRNAs, kinase-dead, or constitutively active SYK mutants. Cotransporter abundance in the plasma membrane was analyzed by biotinylation of cell surface proteins. Results and Discussions: Here we describe that tyrosine phosphorylation of NKCC2 and KCC3 regulates their plasma membrane expression levels. We identified that SYK phosphorylates a specific N-terminal tyrosine residue in each cotransporter. Experimental depletion of endogenous SYK or pharmacological inhibition of its kinase activity increased the abundance of NKCC2 at the plasma membrane of human embryonic kidney cells. In contrast, overexpression of a constitutively active SYK mutant decreased NKCC2 membrane abundance. Intriguingly, the same experimental approaches revealed the opposite effect on KCC3 abundance at the plasma membrane, compatible with the known antagonistic roles of NKCC and KCC cotransporters in cell volume regulation. Conclusion: We identified a novel pathway modulating the cell surface expression of NKCC2 and KCC3 and show that this same pathway has opposite functional outcomes for these two cotransporters. The findings add knowledge on how tumor cells may respond to microenvironmental changes that affect their cell volume or metabolic crosstalk.pt_PT
dc.description.sponsorshipThis work was supported by FCT through a center grant UID/MULTI/04046/2019 to BioISI and PTDC/SAU-ORG/119782/2010 and PTDC/BIA-CEL/28408/2017 to PJ. CAL was recipient of fellowship SFRH/BD/52488/2014 from BioSYS PhD programme PD65-2012 and PB of fellowship SFRH/BPD/94322/2013.pt_PT
dc.description.versionN/Apt_PT
dc.identifier.urihttp://hdl.handle.net/10400.18/7433
dc.language.isoengpt_PT
dc.peerreviewednopt_PT
dc.relationSFRH/BD/52488/2014pt_PT
dc.relationBiosystems & Integrative Sciences Institute
dc.subjectChloride Transportpt_PT
dc.subjectProtein Phosphorylationpt_PT
dc.subjectSYKpt_PT
dc.subjectNKCC2pt_PT
dc.subjectKCC3pt_PT
dc.subjectVias de Transdução de Sinal e Patologias Associadaspt_PT
dc.titleMetabolic tumor cell adaptation: tyrosine phosphorylation modulates cell surface expression of NKCC2 and KCC3pt_PT
dc.typeconference object
dspace.entity.typePublication
oaire.awardTitleBiosystems & Integrative Sciences Institute
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FMulti%2F04046%2F2019/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FSAU-ORG%2F119782%2F2010/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBIA-CEL%2F28408%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBPD%2F94322%2F2013/PT
oaire.citation.conferencePlaceLisboa, Portugalpt_PT
oaire.citation.titleEACR-AACR-ASPIC conference on Tumor Microenvironment, 2-4 March 2020pt_PT
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream3599-PPCDT
oaire.fundingStream3599-PPCDT
oaire.fundingStreamSFRH
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typeconferenceObjectpt_PT
relation.isProjectOfPublication35168786-8dfc-4a00-9759-dab3669fe1ae
relation.isProjectOfPublication4c50eaae-97f4-4471-a751-b4a8db9e3c5d
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relation.isProjectOfPublication.latestForDiscovery710163b5-23b3-4080-82a2-3b5d7a1ff579

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