title: With no lysine L-WNK1 isoforms are negative regulators of the K+-Cl- cotransporters.
abstract: The K(+)-Cl(-) cotransporters (KCC1-KCC4) encompass a branch of the SLC12 family of electroneutral cation-coupled chloride cotransporters that translocate ions out of the cell to regulate various factors, including cell volume and intracellular chloride concentration, among others. L-WNK1 is an ubiquitously expressed kinase that is activated in response to osmotic stress and intracellular chloride depletion, and it is implicated in two distinct hereditary syndromes: the renal disease pseudohypoaldosteronism type II (PHAII) and the neurological disease hereditary sensory neuropathy 2 (HSN2). The effect of L-WNK1 on KCC activity is unknown. Using Xenopus laevis oocytes and HEK-293 cells, we show that the activation of KCCs by cell swelling was prevented by L-WNK1 coexpression. In contrast, the activity of the Na(+)-K(+)-2Cl(-) cotransporter NKCC1 was remarkably increased with L-WNK1 coexpression. The negative effect of L-WNK1 on the KCCs is kinase dependent. Elimination of the STE20 proline-alanine rich kinase (SPAK)/oxidative stress-responsive kinase (OSR1) binding site or the HQ motif required for the WNK-WNK interaction prevented the effect of L-WNK1 on KCCs, suggesting a required interaction between L-WNK1 molecules and SPAK. Together, our data support that NKCC1 and KCCs are coordinately regulated by L-WNK1 isoforms.
authors.affiliation: Department of Nephrology, Instituto Nacional de Cardiología Ignacio Chávez, Tlalpan, Mexico City, Mexico;
authors.forename: Adriana
authors.initials: A
authors.lastname: Mercado
authors.affiliation: División de Investigación, Facultad de Medicina, Universidad Nacional Autónoma de México, Coyoacán, Mexico City, Mexico;
authors.forename: Paola
authors.initials: P
authors.lastname: de Los Heros
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tlalpan, Mexico City, Mexico;
authors.forename: Zesergio
authors.initials: Z
authors.lastname: Melo
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tlalpan, Mexico City, Mexico; Department of Nephrology and Mineral Metabolism, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Tlalpan, Mexico City, Mexico; INSERM UMR970-Paris Cardiovascular Research Center, Paris, France; and University Paris-Descartes, Sorbonne Paris Cité, Faculty of Medicine, Paris, France.
authors.forename: María
authors.initials: M
authors.lastname: Chávez-Canales
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tlalpan, Mexico City, Mexico;
authors.forename: Adrián R
authors.initials: AR
authors.lastname: Murillo-de-Ozores
authors.affiliation: Department of Nephrology and Mineral Metabolism, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Tlalpan, Mexico City, Mexico;
authors.forename: Erika
authors.initials: E
authors.lastname: Moreno
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tlalpan, Mexico City, Mexico; Department of Nephrology and Mineral Metabolism, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Tlalpan, Mexico City, Mexico;
authors.forename: Silvana
authors.initials: S
authors.lastname: Bazúa-Valenti
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tlalpan, Mexico City, Mexico; Department of Nephrology and Mineral Metabolism, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Tlalpan, Mexico City, Mexico;
authors.forename: Norma
authors.initials: N
authors.lastname: Vázquez
authors.affiliation: INSERM UMR970-Paris Cardiovascular Research Center, Paris, France; and University Paris-Descartes, Sorbonne Paris Cité, Faculty of Medicine, Paris, France.
authors.forename: Juliette
authors.initials: J
authors.lastname: Hadchouel
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Tlalpan, Mexico City, Mexico; Department of Nephrology and Mineral Metabolism, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán. Tlalpan, Mexico City, Mexico; gamba@biomedicas.unam.mx.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Gamba
mesh_terms: D000818:Animals; D048429:Cell Size; D057809:HEK293 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008239:Lysine; D015778:Minor Histocompatibility Antigens / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D064587:Osmoregulation; D010766:Phosphorylation; D054730:Protein Interaction Domains and Motifs; D020033:Protein Isoforms; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D064506:Solute Carrier Family 12, Member 2 / Q000378:metabolism*; D027981:Symporters / Q000378:metabolism*; D014162:Transfection; D000076204:WNK Lysine-Deficient Protein Kinase 1; D014982:Xenopus laevis; D000096922:K Cl- Cotransporters
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008239:Lysine; D015778:Minor Histocompatibility Antigens; D020033:Protein Isoforms; D017346:Protein Serine-Threonine Kinases; D064506:Solute Carrier Family 12, Member 2; D027981:Symporters; D000076204:WNK Lysine-Deficient Protein Kinase 1; D000096922:K Cl- Cotransporters; C578877:SLC12A2 protein, human; C118135:OXSR1 protein, human; C486409:STK39 protein, human; C498743:WNK1 protein, human
keywords: HEK-293 cells; SPAK; WNK4; Xenopus oocytes