title: Molecular pathogenesis of inherited hypertension with hyperkalemia: the Na-Cl cotransporter is inhibited by wild-type but not mutant WNK4.
abstract: Mutations in the serine-threonine kinases WNK1 and WNK4 [with no lysine (K) at a key catalytic residue] cause pseudohypoaldosteronism type II (PHAII), a Mendelian disease featuring hypertension, hyperkalemia, hyperchloremia, and metabolic acidosis. Both kinases are expressed in the distal nephron, although the regulators and targets of WNK signaling cascades are unknown. The Cl(-) dependence of PHAII phenotypes, their sensitivity to thiazide diuretics, and the observation that they constitute a "mirror image" of the phenotypes resulting from loss of function mutations in the thiazide-sensitive Na-Cl cotransporter (NCCT) suggest that PHAII may result from increased NCCT activity due to altered WNK signaling. To address this possibility, we measured NCCT-mediated Na(+) influx and membrane expression in the presence of wild-type and mutant WNK4 by heterologous expression in Xenopus oocytes. Wild-type WNK4 inhibits NCCT-mediated Na-influx by reducing membrane expression of the cotransporter ((22)Na-influx reduced 50%, P < 1 x 10(-9), surface expression reduced 75%, P < 1 x 10(-14) in the presence of WNK4). This inhibition depends on WNK4 kinase activity, because missense mutations that abrogate kinase function prevent this effect. PHAII-causing missense mutations, which are remote from the kinase domain, also prevent inhibition of NCCT activity, providing insight into the pathophysiology of the disorder. The specificity of this effect is indicated by the finding that WNK4 and the carboxyl terminus of NCCT coimmunoprecipitate when expressed in HEK 293T cells. Together, these findings demonstrate that WNK4 negatively regulates surface expression of NCCT and implicate loss of this regulation in the molecular pathogenesis of an inherited form of hypertension.
authors.affiliation: Department of Genetics, Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, CT 06510, USA.
authors.forename: Frederick H
authors.initials: FH
authors.lastname: Wilson
authors.forename: Kristopher T
authors.initials: KT
authors.lastname: Kahle
authors.forename: Ernesto
authors.initials: E
authors.lastname: Sabath
authors.forename: Maria D
authors.initials: MD
authors.lastname: Lalioti
authors.forename: Alicia K
authors.initials: AK
authors.lastname: Rapson
authors.forename: Robert S
authors.initials: RS
authors.lastname: Hoover
authors.forename: Steven C
authors.initials: SC
authors.lastname: Hebert
authors.forename: Gerardo
authors.initials: G
authors.lastname: Gamba
authors.forename: Richard P
authors.initials: RP
authors.lastname: Lifton
mesh_terms: D000818:Animals; D001483:Base Sequence; D005260:Female; D006947:Hyperkalemia / Q000235:genetics*; D006973:Hypertension / Q000235:genetics*; D017136:Ion Transport; D051379:Mice; D008969:Molecular Sequence Data; D017346:Protein Serine-Threonine Kinases / Q000494:pharmacology / Q000502:physiology*; D012964:Sodium / Q000378:metabolism; D049992:Sodium Chloride Symporters; D027981:Symporters / Q000037:antagonists & inhibitors* / Q000502:physiology; D029867:Xenopus Proteins / Q000494:pharmacology*; D014982:Xenopus laevis
chemical_list: D049992:Sodium Chloride Symporters; D027981:Symporters; D029867:Xenopus Proteins; D012964:Sodium; C499024:Prkwnk4 protein, mouse; D017346:Protein Serine-Threonine Kinases; C519693:WNK4 protein, Xenopus