Activation of the thiazide-sensitive Na+-Cl- cotransporter by the WNK-regulated kinases SPAK and OSR1

Ciaran Richardson, Fatema H. Rafiqi, Hakan K. R. Karlsson, Ntsane Moleleki, Alain Vandewalle, David G. Campbell, Nick A. Morrice, Dario R. Alessi

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    254 Citations (Scopus)

    Abstract

    Mutations increasing WNK1 kinase expression in humans cause the pseudohypoaldosteronism type II hypertension syndrome. This condition is treated effectively by thiazide diuretics, which exert their effects by inhibiting the Na+-Cl- cotransporter (NCC), suggesting a link between WNK1 and NCC. Here, we demonstrate that the SPAK and OSR1 kinases that are activated by WNK1 phosphorylate human NCC at three conserved residues (Thr46, Thr55 and Thr60). Activation of the WNK1-SPAK/OSR1 signalling pathway by treatment of HEK293 or mpkDCT kidney distal-convoluted-tubule-derived cells with hypotonic low-chloride conditions induced phosphorylation of NCC at residues phosphorylated by SPAK/OSR1. Efficient phosphorylation of NCC was dependent upon a docking interaction between an RFXI motif in NCC and SPAK/OSR1. Mutation of Thr60 to Ala in NCC markedly inhibited phosphorylation of Thr46 and Thr55 as well as NCC activation induced by hypotonic low-chloride treatment of HEK293 cells. Our results establish that the WNK1-SPAK/OSR1 signalling pathway plays a key role in controlling the phosphorylation and activity of NCC. They also suggest a mechanism by which increased WNK1 overexpression could lead to hypertension and that inhibitors of SPAK/OSR1 might be of use in reducing blood pressure by suppressing phosphorylation and hence activity of NCC.

    Original languageEnglish
    Pages (from-to)675-684
    Number of pages10
    JournalJournal of Cell Science
    Volume121
    Issue number5
    Early online date12 Feb 2008
    DOIs
    Publication statusPublished - 1 Mar 2008

    Keywords

    • Cell signalling
    • Kinase
    • OSR1
    • SPAK
    • WNK1
    • Cotransporter

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