WNK3 abrogates the NEDD4-2-mediated inhibition of the renal Na+-Cl- cotransporter.

Détails

ID Serval
serval:BIB_46E058E53C8F
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
WNK3 abrogates the NEDD4-2-mediated inhibition of the renal Na+-Cl- cotransporter.
Périodique
American Journal of Physiology. Renal Physiology
Auteur(s)
Lagnaz D., Arroyo J.P., Chávez-Canales M., Vázquez N., Rizzo F., Spirlí A., Debonneville A., Staub O., Gamba G.
ISSN
1522-1466 (Electronic)
ISSN-L
1522-1466
Statut éditorial
Publié
Date de publication
2014
Peer-reviewed
Oui
Volume
307
Numéro
3
Pages
F275-F286
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Résumé
The serine/threonine kinase WNK3 and the ubiquitin-protein ligase NEDD4-2 are key regulators of the thiazide-sensitive Na+-Cl- cotransporter (NCC), WNK3 as an activator and NEDD2-4 as an inhibitor. Nedd4-2 was identified as an interacting partner of WNK3 through a glutathione-S-transferase pull-down assay using the N-terminal domain of WNK3, combined with LC-MS/MS analysis. This was validated by coimmunoprecipitation of WNK3 and NEDD4-2 expressed in HEK293 cells. Our data also revealed that the interaction between Nedd4-2 and WNK3 does not involve the PY-like motif found in WNK3. The level of WNK3 ubiquitylation did not change when NEDD4-2 was expressed in HEK293 cells. Moreover, in contrast to SGK1, WNK3 did not phosphorylate NEDD4-2 on S222 or S328. Coimmunoprecipitation assays showed that WNK3 does not regulate the interaction between NCC and NEDD4-2. Interestingly, in Xenopus laevis oocytes, WNK3 was able to recover the SGK1-resistant NEDD4-2 S222A/S328A-mediated inhibition of NCC and further activate NCC. Furthermore, elimination of the SPAK binding site in the kinase domain of WNK3 (WNK3-F242A, which lacks the capacity to bind the serine/threonine kinase SPAK) prevented the WNK3 NCC-activating effect, but not the Nedd4-2-inhibitory effect. Together, these results suggest that a novel role for WNK3 on NCC expression at the plasma membrane, an effect apparently independent of the SPAK kinase and the aldosterone-SGK1 pathway.
Pubmed
Web of science
Création de la notice
25/09/2014 17:57
Dernière modification de la notice
03/03/2018 16:48
Données d'usage