WNK4 inhibits plasma membrane targeting of NCC through regulation of syntaxin13 SNARE formation

Woo Young Chung, Hyun Woo Park, Jung Woo Han, Min Goo Lee, Joo Young Kim

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

WNK4, a serine/threonine kinase, plays a critical role in the expression of membrane proteins in the cell surface; however, the underlying mechanism of WNK4 is not clear. Here, we demonstrate that WNK4 inhibits the fusion of plasma membrane delivering vesicle with sorting/recycling endosome through disrupting SNARE formation of syntaxin13, an endosomal t-SNARE and VAMP2, the v-SNARE in plasma membrane delivering vesicle. Their interaction and co-localization were enhanced by hyperosmotic stimulation which is known for WNK4 activation. The kinase domain of WNK4 interacts with the transmembrane domain (TM) of syntaxin13 and this interaction was abolished when the TM was replaced with that of syntaxin16. Interestingly, cell fractionation using sucrose gradients revealed that WNK4 inhibited the formation of the syntaxin13/VAMP2 SNARE complex in the endosomal compartment, but not syntaxin16/VAMP2 or syntaxin13/VAMP7. Syntaxin13 was not phosphorylated by WNK4 and WNK4KI also showed the same binding strength and similar inhibitory regulation on SNARE formation of syntaxin13. Physiological relevance of this mechanism was proved with the expression of NCC (Na+ C1- co-transporter) in the cell surface. The inhibiting activity of WNK4 on surface expression of NCC was abolished by syntaxin13 siRNA transfection. These results suggest that WNK4 attenuates PM targeting of NCC proteins through regulation of syntaxin13 SNARE complex formation with VAMP2 in recycling and sorting endosome.

Original languageEnglish
Pages (from-to)2469-2477
Number of pages9
JournalCellular Signalling
Volume25
Issue number12
DOIs
Publication statusPublished - 2013 Jan 1

Fingerprint

Symporters
SNARE Proteins
Vesicle-Associated Membrane Protein 2
Cell Membrane
Endosomes
Membrane Proteins
Cell Fractionation
Protein-Serine-Threonine Kinases
Recycling
Small Interfering RNA
Transfection
Sucrose
Phosphotransferases
Proteins

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Chung, Woo Young ; Park, Hyun Woo ; Han, Jung Woo ; Lee, Min Goo ; Kim, Joo Young. / WNK4 inhibits plasma membrane targeting of NCC through regulation of syntaxin13 SNARE formation. In: Cellular Signalling. 2013 ; Vol. 25, No. 12. pp. 2469-2477.
@article{c733c808e65a4ed2af15a740b7dcad9d,
title = "WNK4 inhibits plasma membrane targeting of NCC through regulation of syntaxin13 SNARE formation",
abstract = "WNK4, a serine/threonine kinase, plays a critical role in the expression of membrane proteins in the cell surface; however, the underlying mechanism of WNK4 is not clear. Here, we demonstrate that WNK4 inhibits the fusion of plasma membrane delivering vesicle with sorting/recycling endosome through disrupting SNARE formation of syntaxin13, an endosomal t-SNARE and VAMP2, the v-SNARE in plasma membrane delivering vesicle. Their interaction and co-localization were enhanced by hyperosmotic stimulation which is known for WNK4 activation. The kinase domain of WNK4 interacts with the transmembrane domain (TM) of syntaxin13 and this interaction was abolished when the TM was replaced with that of syntaxin16. Interestingly, cell fractionation using sucrose gradients revealed that WNK4 inhibited the formation of the syntaxin13/VAMP2 SNARE complex in the endosomal compartment, but not syntaxin16/VAMP2 or syntaxin13/VAMP7. Syntaxin13 was not phosphorylated by WNK4 and WNK4KI also showed the same binding strength and similar inhibitory regulation on SNARE formation of syntaxin13. Physiological relevance of this mechanism was proved with the expression of NCC (Na+ C1- co-transporter) in the cell surface. The inhibiting activity of WNK4 on surface expression of NCC was abolished by syntaxin13 siRNA transfection. These results suggest that WNK4 attenuates PM targeting of NCC proteins through regulation of syntaxin13 SNARE complex formation with VAMP2 in recycling and sorting endosome.",
author = "Chung, {Woo Young} and Park, {Hyun Woo} and Han, {Jung Woo} and Lee, {Min Goo} and Kim, {Joo Young}",
year = "2013",
month = "1",
day = "1",
doi = "10.1016/j.cellsig.2013.08.006",
language = "English",
volume = "25",
pages = "2469--2477",
journal = "Cellular Signalling",
issn = "0898-6568",
publisher = "Elsevier Inc.",
number = "12",

}

WNK4 inhibits plasma membrane targeting of NCC through regulation of syntaxin13 SNARE formation. / Chung, Woo Young; Park, Hyun Woo; Han, Jung Woo; Lee, Min Goo; Kim, Joo Young.

In: Cellular Signalling, Vol. 25, No. 12, 01.01.2013, p. 2469-2477.

Research output: Contribution to journalArticle

TY - JOUR

T1 - WNK4 inhibits plasma membrane targeting of NCC through regulation of syntaxin13 SNARE formation

AU - Chung, Woo Young

AU - Park, Hyun Woo

AU - Han, Jung Woo

AU - Lee, Min Goo

AU - Kim, Joo Young

PY - 2013/1/1

Y1 - 2013/1/1

N2 - WNK4, a serine/threonine kinase, plays a critical role in the expression of membrane proteins in the cell surface; however, the underlying mechanism of WNK4 is not clear. Here, we demonstrate that WNK4 inhibits the fusion of plasma membrane delivering vesicle with sorting/recycling endosome through disrupting SNARE formation of syntaxin13, an endosomal t-SNARE and VAMP2, the v-SNARE in plasma membrane delivering vesicle. Their interaction and co-localization were enhanced by hyperosmotic stimulation which is known for WNK4 activation. The kinase domain of WNK4 interacts with the transmembrane domain (TM) of syntaxin13 and this interaction was abolished when the TM was replaced with that of syntaxin16. Interestingly, cell fractionation using sucrose gradients revealed that WNK4 inhibited the formation of the syntaxin13/VAMP2 SNARE complex in the endosomal compartment, but not syntaxin16/VAMP2 or syntaxin13/VAMP7. Syntaxin13 was not phosphorylated by WNK4 and WNK4KI also showed the same binding strength and similar inhibitory regulation on SNARE formation of syntaxin13. Physiological relevance of this mechanism was proved with the expression of NCC (Na+ C1- co-transporter) in the cell surface. The inhibiting activity of WNK4 on surface expression of NCC was abolished by syntaxin13 siRNA transfection. These results suggest that WNK4 attenuates PM targeting of NCC proteins through regulation of syntaxin13 SNARE complex formation with VAMP2 in recycling and sorting endosome.

AB - WNK4, a serine/threonine kinase, plays a critical role in the expression of membrane proteins in the cell surface; however, the underlying mechanism of WNK4 is not clear. Here, we demonstrate that WNK4 inhibits the fusion of plasma membrane delivering vesicle with sorting/recycling endosome through disrupting SNARE formation of syntaxin13, an endosomal t-SNARE and VAMP2, the v-SNARE in plasma membrane delivering vesicle. Their interaction and co-localization were enhanced by hyperosmotic stimulation which is known for WNK4 activation. The kinase domain of WNK4 interacts with the transmembrane domain (TM) of syntaxin13 and this interaction was abolished when the TM was replaced with that of syntaxin16. Interestingly, cell fractionation using sucrose gradients revealed that WNK4 inhibited the formation of the syntaxin13/VAMP2 SNARE complex in the endosomal compartment, but not syntaxin16/VAMP2 or syntaxin13/VAMP7. Syntaxin13 was not phosphorylated by WNK4 and WNK4KI also showed the same binding strength and similar inhibitory regulation on SNARE formation of syntaxin13. Physiological relevance of this mechanism was proved with the expression of NCC (Na+ C1- co-transporter) in the cell surface. The inhibiting activity of WNK4 on surface expression of NCC was abolished by syntaxin13 siRNA transfection. These results suggest that WNK4 attenuates PM targeting of NCC proteins through regulation of syntaxin13 SNARE complex formation with VAMP2 in recycling and sorting endosome.

UR - http://www.scopus.com/inward/record.url?scp=84884142912&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84884142912&partnerID=8YFLogxK

U2 - 10.1016/j.cellsig.2013.08.006

DO - 10.1016/j.cellsig.2013.08.006

M3 - Article

VL - 25

SP - 2469

EP - 2477

JO - Cellular Signalling

JF - Cellular Signalling

SN - 0898-6568

IS - 12

ER -