title: WNK kinases influence TRPV4 channel function and localization.
abstract: TRPV4, a renally expressed nonselective cation channel of the transient receptor potential (TRP) family, is gated by hypotonicity. Kinases of the WNK family influence expression and function of the thiazide-sensitive Na+-Cl- cotransporter, and monogenic human hypertension has been linked to mutations in the gene coding for WNK4. Along with TRPV4, WNK isoforms are highly expressed in the distal nephron. We show here that coexpression of WNK4 downregulates TRPV4 function in human embryonic kidney (HEK-293) cells and that this effect is mediated via decreased cell surface expression of TRPV4; total abundance of TRPV4 in whole cell lysates is unaffected. The effect of the related kinase WNK1 on TRPV4 function and surface expression was similar to that of WNK4. Disease-causing point mutations in WNK4 abrogate, but do not eliminate, the inhibitory effect on TRPV4 function. In contrast to wild-type WNK4, a kinase-dead WNK4 point mutant failed to influence TRPV4 trafficking; however, deletion of the entire WNK4 kinase domain did not blunt the effect of WNK4 on localization of TRPV4. Deletion of the extreme COOH-terminal putative coiled-coil domain of WNK4 abolished its effect. In immunoprecipitation experiments, we were unable to detect direct interaction between TRPV4 and either WNK kinase. In aggregate, these data indicate that TRPV4 is functionally regulated by WNK family kinases at the level of cell surface expression. Because TRPV4 and WNK kinases are coexpressed in the distal nephron in vivo and because there is a tendency toward hypercalcemia in TRPV4-/- mice, we speculate that this pathway may impact systemic Ca2+ balance. In addition, because WNK kinases and TRPV4 are activated by anisotonicity, they may comprise elements of an osmosensing or osmotically responsive signal transduction cascade in the distal nephron.
authors.affiliation: Department of Medicine, Oregon Health & Science Univ., Portland, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Fu
authors.forename: Arohan
authors.initials: A
authors.lastname: Subramanya
authors.forename: David
authors.initials: D
authors.lastname: Rozansky
authors.forename: David M
authors.initials: DM
authors.lastname: Cohen
mesh_terms: D000818:Animals; D019921:Biotinylation; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D004622:Embryo, Mammalian; D017353:Gene Deletion; D006706:Homeostasis; D006801:Humans; D006973:Hypertension / Q000235:genetics; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins; D007668:Kidney; D051379:Mice; D018345:Mice, Knockout; D015778:Minor Histocompatibility Antigens; D009154:Mutation; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology*; D051381:Rats; D011994:Recombinant Proteins; D050916:TRPV Cation Channels / Q000032:analysis* / Q000172:deficiency / Q000502:physiology*; D014162:Transfection; D000076204:WNK Lysine-Deficient Protein Kinase 1
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D015778:Minor Histocompatibility Antigens; D011994:Recombinant Proteins; D050916:TRPV Cation Channels; C416986:TRPV4 protein, human; C486576:Trpv4 protein, mouse; C499024:Prkwnk4 protein, mouse; D017346:Protein Serine-Threonine Kinases; D000076204:WNK Lysine-Deficient Protein Kinase 1; C498743:WNK1 protein, human; C433806:WNK4 protein, human; C410410:Wnk1 protein, rat; D002118:Calcium