PXD016218

PXD016218 is an original dataset announced via ProteomeXchange.

Title	WNK1-specific phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1
Description	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 case of insulin-responsive cells, which activate protein kinase AKT to phosphorylate TBC1D4, a RAB-GAP 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-specific 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 or therapeutic options in type 2 diabetes.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_05:29:57.620.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Rune Matthiesen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2019-11-11 02:36:18	ID requested
1	2021-11-03 02:40:40	announced
⏵ 2	2024-10-22 05:29:58	announced	2024-10-22: Updated project metadata.

10.1016/j.abb.2019.108223;

Henriques AFA, Matos P, Carvalho AS, Azkargorta M, Elortza F, Matthiesen R, Jordan P, WNK1 phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1. Arch Biochem Biophys, 679():108223(2020) [pubmed]

submitter keyword: Human, phosphorylation

Rune Matthiesen
contact affiliation	CEDOC, NMS, Lisbon, Portugal
contact email	runem2009@gmail.com
lab head
Rune Matthiesen
contact affiliation	Computational and Experimental Biology Group, CEDOC NOVA MEDICAL SCHOOL / FACULDADE DE CIÊNCIAS MÉDICAS UNIVERSIDADE NOVA DE LISBOA Rua Câmara Pestana, 6-6A | 1150-082 Lisboa Portugal
contact email	runem2009@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD016218
     1. Label: PRIDE project
     2. Name: WNK1-specific phosphorylation sites in TBC1D1 and TBC1D4 modulate cell surface expression of GLUT1