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Proc Natl Acad Sci U S A
2008 Jun 17;10524:8458-63. doi: 10.1073/pnas.0802966105.
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Regulation of NKCC2 by a chloride-sensing mechanism involving the WNK3 and SPAK kinases.
Ponce-Coria J, San-Cristobal P, Kahle KT, Vazquez N, Pacheco-Alvarez D, de Los Heros P, Juárez P, Muñoz E, Michel G, Bobadilla NA, Gimenez I, Lifton RP, Hebert SC, Gamba G.
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The Na(+):K(+):2Cl(-) cotransporter (NKCC2) is the target of loop diuretics and is mutated in Bartter's syndrome, a heterogeneous autosomal recessive disease that impairs salt reabsorption in the kidney's thick ascending limb (TAL). Despite the importance of this cation/chloride cotransporter (CCC), the mechanisms that underlie its regulation are largely unknown. Here, we show that intracellular chloride depletion in Xenopus laevis oocytes, achieved by either coexpression of the K-Cl cotransporter KCC2 or low-chloride hypotonic stress, activates NKCC2 by promoting the phosphorylation of three highly conserved threonines (96, 101, and 111) in the amino terminus. Elimination of these residues renders NKCC2 unresponsive to reductions of [Cl(-)](i). The chloride-sensitive activation of NKCC2 requires the interaction of two serine-threonine kinases, WNK3 (related to WNK1 and WNK4, genes mutated in a Mendelian form of hypertension) and SPAK (a Ste20-type kinase known to interact with and phosphorylate other CCCs). WNK3 is positioned upstream of SPAK and appears to be the chloride-sensitive kinase. Elimination of WNK3's unique SPAK-binding motif prevents its activation of NKCC2, as does the mutation of threonines 96, 101, and 111. A catalytically inactive WNK3 mutant also completely prevents NKCC2 activation by intracellular chloride depletion. Together these data reveal a chloride-sensing mechanism that regulates NKCC2 and provide insight into how increases in the level of intracellular chloride in TAL cells, as seen in certain pathological states, could drastically impair renal salt reabsorption.
Ackerman,
Hypotonicity activates a native chloride current in Xenopus oocytes.
1994, Pubmed,
Xenbase
Ackerman,
Hypotonicity activates a native chloride current in Xenopus oocytes.
1994,
Pubmed
,
Xenbase Adragna,
Regulation of K-Cl cotransport: from function to genes.
2004,
Pubmed Darman,
A regulatory locus of phosphorylation in the N terminus of the Na-K-Cl cotransporter, NKCC1.
2002,
Pubmed Darman,
Modulation of ion transport by direct targeting of protein phosphatase type 1 to the Na-K-Cl cotransporter.
2001,
Pubmed de Los Heros,
WNK3 bypasses the tonicity requirement for K-Cl cotransporter activation via a phosphatase-dependent pathway.
2006,
Pubmed
,
Xenbase Delpire,
Genome-wide analysis of SPAK/OSR1 binding motifs.
2007,
Pubmed Delpire,
SPAK and OSR1, key kinases involved in the regulation of chloride transport.
2006,
Pubmed Dowd,
PASK (proline-alanine-rich STE20-related kinase), a regulatory kinase of the Na-K-Cl cotransporter (NKCC1).
2003,
Pubmed Flatman,
Cotransporters, WNKs and hypertension: important leads from the study of monogenetic disorders of blood pressure regulation.
2007,
Pubmed Flemmer,
Activation of the Na-K-Cl cotransporter NKCC1 detected with a phospho-specific antibody.
2002,
Pubmed Gagnon,
Characterization of SPAK and OSR1, regulatory kinases of the Na-K-2Cl cotransporter.
2006,
Pubmed
,
Xenbase Gagnon,
Volume sensitivity of cation-Cl- cotransporters is modulated by the interaction of two kinases: Ste20-related proline-alanine-rich kinase and WNK4.
2006,
Pubmed
,
Xenbase Gamba,
Molecular biology of distal nephron sodium transport mechanisms.
1999,
Pubmed Gamba,
Molecular physiology and pathophysiology of electroneutral cation-chloride cotransporters.
2005,
Pubmed Gamba,
Molecular cloning, primary structure, and characterization of two members of the mammalian electroneutral sodium-(potassium)-chloride cotransporter family expressed in kidney.
1994,
Pubmed
,
Xenbase Garzón-Muvdi,
WNK4 kinase is a negative regulator of K+-Cl- cotransporters.
2007,
Pubmed
,
Xenbase Giménez,
Regulatory phosphorylation sites in the NH2 terminus of the renal Na-K-Cl cotransporter (NKCC2).
2005,
Pubmed
,
Xenbase Giménez,
Short-term stimulation of the renal Na-K-Cl cotransporter (NKCC2) by vasopressin involves phosphorylation and membrane translocation of the protein.
2003,
Pubmed Hebert,
Bartter syndrome.
2003,
Pubmed Horster,
Embryonic epithelial membrane transporters.
2000,
Pubmed Kahle,
WNK3 modulates transport of Cl- in and out of cells: implications for control of cell volume and neuronal excitability.
2005,
Pubmed
,
Xenbase Keicher,
Endogenous Na(+)-K+ (or NH4+)-2Cl- cotransport in Rana oocytes; anomalous effect of external NH4+ on pHi.
1994,
Pubmed Lenertz,
Properties of WNK1 and implications for other family members.
2005,
Pubmed Lytle,
Coordinate modulation of Na-K-2Cl cotransport and K-Cl cotransport by cell volume and chloride.
2002,
Pubmed Lytle,
Regulatory phosphorylation of the secretory Na-K-Cl cotransporter: modulation by cytoplasmic Cl.
1996,
Pubmed Lytle,
The Na-K-Cl cotransport protein of shark rectal gland. II. Regulation by direct phosphorylation.
1992,
Pubmed Meade,
cAMP-dependent activation of the renal-specific Na+-K+-2Cl- cotransporter is mediated by regulation of cotransporter trafficking.
2003,
Pubmed
,
Xenbase Mercado,
Functional comparison of the K+-Cl- cotransporters KCC1 and KCC4.
2000,
Pubmed
,
Xenbase Moriguchi,
WNK1 regulates phosphorylation of cation-chloride-coupled cotransporters via the STE20-related kinases, SPAK and OSR1.
2005,
Pubmed Mount,
Cloning and characterization of KCC3 and KCC4, new members of the cation-chloride cotransporter gene family.
1999,
Pubmed
,
Xenbase Pacheco-Alvarez,
The Na+:Cl- cotransporter is activated and phosphorylated at the amino-terminal domain upon intracellular chloride depletion.
2006,
Pubmed
,
Xenbase Paredes,
Activity of the renal Na+-K+-2Cl- cotransporter is reduced by mutagenesis of N-glycosylation sites: role for protein surface charge in Cl- transport.
2006,
Pubmed
,
Xenbase Piechotta,
Cation chloride cotransporters interact with the stress-related kinases Ste20-related proline-alanine-rich kinase (SPAK) and oxidative stress response 1 (OSR1).
2002,
Pubmed Piechotta,
Characterization of the interaction of the stress kinase SPAK with the Na+-K+-2Cl- cotransporter in the nervous system: evidence for a scaffolding role of the kinase.
2003,
Pubmed
,
Xenbase Plata,
Functional properties of the apical Na+-K+-2Cl- cotransporter isoforms.
2002,
Pubmed
,
Xenbase Rinehart,
WNK3 kinase is a positive regulator of NKCC2 and NCC, renal cation-Cl- cotransporters required for normal blood pressure homeostasis.
2005,
Pubmed
,
Xenbase Simon,
Bartter's syndrome, hypokalaemic alkalosis with hypercalciuria, is caused by mutations in the Na-K-2Cl cotransporter NKCC2.
1996,
Pubmed Song,
Molecular, functional, and genomic characterization of human KCC2, the neuronal K-Cl cotransporter.
2002,
Pubmed
,
Xenbase Vitari,
The WNK1 and WNK4 protein kinases that are mutated in Gordon's hypertension syndrome phosphorylate and activate SPAK and OSR1 protein kinases.
2005,
Pubmed Vitari,
Functional interactions of the SPAK/OSR1 kinases with their upstream activator WNK1 and downstream substrate NKCC1.
2006,
Pubmed Xu,
WNK1, a novel mammalian serine/threonine protein kinase lacking the catalytic lysine in subdomain II.
2000,
Pubmed
