WNK4 is indispensable for the pathogenesis of pseudohypoaldosteronism type II caused by mutant KLHL3

Biochem Biophys Res Commun. 2017 Sep 23;491(3):727-732. doi: 10.1016/j.bbrc.2017.07.121. Epub 2017 Jul 22.


WNK-OSR1/SPAK-NCC signaling cascade is important for regulating salt balance and blood pressure. Activation of WNK-OSR1/SPAK-NaCl cotransporter (NCC) cascade increases sodium reabsorption in the kidney, leading to pseudohypoaldosteronism type II (PHA II) characterized by salt-sensitive hypertension and hyperkalemia. It has been previously demonstrated that the amount of phosphorylated and total NCC markedly decreased in WNK4-/- mice, indicating that WNK4 plays a major role for activation of OSR1/SPAK-NCC signaling. However, it is unclear whether absence of WNK4 can be compensated by other WNK kinases. We recently reported that KLHL3R528H/+ knock-in mice, a PHAII model, exhibited augmented activation of OSR1/SPAK-NCC signaling by increased protein levels of both WNK1 and WNK4 due to impaired protein degradation by the mutant KLHL3. In this study, we sought to determine the contribution of WNK4 to OSR1/SPAK-NCC signaling using an in vivo model which shows extremely increased WNK1 with absence of WNK4. We generated WNK4-/-KLHL3R528H/+ mice and WNK4-/-KLHL3R528H/R528H mice by crossing WNK4-/- mice with KLHL3R528H/+ mice. Thereafter, WNK-OSR1/SPAK-NCC phosphorylation signal cascade was examined in kidneys from these mice. As expected, both WNK4-/-KLHL3R528H/+ mice and WNK4-/-KLHL3R528H/R528H mice demonstrated increased WNK1 in the kidney, due to the KLHL3 mutation, and WNK4 deficiency. However, phosphorylation of SPAK and NCC at distal convoluted tubules were almost completely absent even in WNK4-/-KLHL3R528H/R528H mice. In conclusion, increased WNK1 was unable to compensate for WNK4 deficiency and phosphorylate the NCC, indicating that WNK4 is indispensable for the onset of PHAII.

Keywords: KLHL3; Pseudohypoaldosteronism type II; Salt-sensitive hypertension; WNK kinase.

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Animals
  • Female
  • Kidney / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Microfilament Proteins / genetics*
  • Microfilament Proteins / metabolism*
  • Minor Histocompatibility Antigens / metabolism*
  • Mutation / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Pseudohypoaldosteronism / genetics*
  • Pseudohypoaldosteronism / metabolism*
  • Structure-Activity Relationship
  • WNK Lysine-Deficient Protein Kinase 1


  • Adaptor Proteins, Signal Transducing
  • KLHL3 protein, mouse
  • Microfilament Proteins
  • Minor Histocompatibility Antigens
  • Prkwnk4 protein, mouse
  • Protein Serine-Threonine Kinases
  • WNK Lysine-Deficient Protein Kinase 1
  • Wnk1 protein, mouse