Kir5.1 regulates Nedd4-2-mediated ubiquitination of Kir4.1 in distal nephron.

Détails

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Etat: Public
Version: Author's accepted manuscript
ID Serval
serval:BIB_177C962852AF
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Kir5.1 regulates Nedd4-2-mediated ubiquitination of Kir4.1 in distal nephron.
Périodique
American journal of physiology. Renal physiology
Auteur(s)
Wang M.X., Su X.T., Wu P., Gao Z.X., Wang W.H., Staub O., Lin D.H.
ISSN
1522-1466 (Electronic)
ISSN-L
1522-1466
Statut éditorial
Publié
Date de publication
01/10/2018
Peer-reviewed
Oui
Volume
315
Numéro
4
Pages
F986-F996
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
Kir4.1/5.1 heterotetramer participates in generating the negative cell membrane potential in distal convoluted tubule (DCT) and plays a critical role in determining the activity of Na-Cl cotransporter (NCC). Kir5.1 contains a phosphothreonine motif at its COOH terminus (AA249-252). Coimmunoprecipitation showed that Nedd4-2 was associated with Kir5.1 in HEK293 cells cotransfected with Kir5.1 or Kir4.1/Kir5.1. GST pull-down further confirmed the association between Nedd4-2 and Kir5.1. Ubiquitination assay showed that Nedd4-2 increased the ubiquitination of Kir4.1/Kir5.1 heterotetramer in the cells cotransfected with Kir4.1/Kir5.1, but it has no effect on Kir4.1 or Kir5.1 alone. Patch-clamp and Western blot also demonstrated that coexpression of Nedd4-2 but not Nedd4-1 decreased K currents and Kir4.1 expression in the cells cotransfected with Kir4.1 and Kir5.1. In contrast, Nedd4-2 fails to inhibit Kir4.1 in the absence of Kir5.1 or in the cells transfected with the inactivated form of Nedd4-2 (Nedd4-2C821A). Moreover, the mutation of TPVT motif in the COOH terminus of Kir5.1 largely abolished the association of Nedd4-2 with Kir5.1 and abolished the inhibitory effect of Nedd4-2 on K currents in HEK293 cells transfected with Kir4.1 and Kir5.1 mutant (Kir5.1T249A). Finally, the basolateral K conductance in the DCT and Kir4.1 expression is significantly increased in the kidney-specific Nedd4-2 knockout or in Kir5.1 knockout mice in comparison to their corresponding wild-type littermates. We conclude that Nedd4-2 binds to Kir5.1 at the phosphothreonine motif of the COOH terminus, and the association of Nedd4-2 with Kir5.1 facilitates the ubiquitination of Kir4.1, thereby regulating its plasma expression in the DCT.
Mots-clé
EAST/SeSAME syndrome, Kcnj10, Kcnj16, NCC, Nedd4-1
Pubmed
Web of science
Création de la notice
25/06/2018 8:52
Dernière modification de la notice
20/08/2019 12:47
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