Kidney-specific WNK1 isoform (KS-WNK1) is a potent activator of WNK4 and NCC

Am J Physiol Renal Physiol. 2018 Sep 1;315(3):F734-F745. doi: 10.1152/ajprenal.00145.2018. Epub 2018 May 30.


Familial hyperkalemic hypertension (FHHt) can be mainly attributed to increased activity of the renal Na+:Cl- cotransporter (NCC), which is caused by altered expression and regulation of the with-no-lysine (K) 1 (WNK1) or WNK4 kinases. The WNK1 gene gives rise to a kidney-specific isoform that lacks the kinase domain (KS-WNK1), the expression of which occurs primarily in the distal convoluted tubule. The role played by KS-WNK1 in the modulation of the WNK/STE20-proline-alanine rich kinase (SPAK)/NCC pathway remains elusive. In the present study, we assessed the effect of human KS-WNK1 on NCC activity and on the WNK4-SPAK pathway. Microinjection of oocytes with human KS-WNK1 cRNA induces remarkable activation and phosphorylation of SPAK and NCC. The effect of KS-WNK1 was abrogated by eliminating a WNK-WNK-interacting domain and by a specific WNK inhibitor, WNK463, indicating that the activation of SPAK/NCC by KS-WNK1 is due to interaction with another WNK kinase. Under control conditions in oocytes, the activating serine 335 of the WNK4 T loop is not phosphorylated. In contrast, this serine becomes phosphorylated when the intracellular chloride concentration ([Cl-]i) is reduced or when KS-WNK1 is coexpressed with WNK4. KS-WNK1-mediated activation of WNK4 is not due to a decrease of the [Cl-]i. Coimmunoprecipitation analysis revealed that KS-WNK1 and WNK4 interact with each other and that WNK4 becomes autophosphorylated at serine 335 when it is associated with KS-WNK1. Together, these observations suggest that WNK4 becomes active in the presence of KS-WNK1, despite a constant [Cl-]i.

Keywords: Na+:Cl− cotransporter; STE20-proline-alanine rich kinase; distal convoluted tubule; diuretics; salt transport.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chlorides / metabolism*
  • Enzyme Activation
  • Female
  • Humans
  • Kidney / enzymology*
  • Oocytes
  • Phosphorylation
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Rats
  • Sodium / metabolism*
  • Solute Carrier Family 12, Member 3 / genetics
  • Solute Carrier Family 12, Member 3 / metabolism
  • WNK Lysine-Deficient Protein Kinase 1 / genetics
  • WNK Lysine-Deficient Protein Kinase 1 / metabolism*
  • Xenopus Proteins / metabolism
  • Xenopus laevis


  • Chlorides
  • Slc12a3 protein, rat
  • Solute Carrier Family 12, Member 3
  • Xenopus Proteins
  • Sodium
  • Protein Serine-Threonine Kinases
  • STK39 protein, Xenopus
  • WNK Lysine-Deficient Protein Kinase 1
  • WNK1 protein, human
  • WNK4 protein, human