title: SPAK and OSR1 play essential roles in potassium homeostasis through actions on the distal convoluted tubule.
abstract: STE20 (Sterile 20)/SPS-1 related proline/alanine-rich kinase (SPAK) and oxidative stress-response kinase-1 (OSR1) phosphorylate and activate the renal Na(+) -K(+) -2Cl(-) cotransporter 2 (NKCC2) and Na(+) Cl(-) cotransporter (NCC). Mouse models suggest that OSR1 mainly activates NKCC2-mediated sodium transport along the thick ascending limb, while SPAK mainly activates NCC along the distal convoluted tubule, but the kinases may compensate for each other. We hypothesized that disruption of both kinases would lead to polyuria and severe salt-wasting, and generated SPAK/OSR1 double knockout mice to test this. Despite a lack of SPAK and OSR1, phosphorylated NKCC2 abundance was still high, suggesting the existence of an alternative activating kinase. Compensatory changes in SPAK/OSR1-independent phosphorylation sites on both NKCC2 and NCC and changes in sodium transport along the collecting duct were also observed. Potassium restriction revealed that SPAK and OSR1 play essential roles in the emerging model that NCC activation is central to sensing changes in plasma [K(+) ].
  
  
  
  STE20 (Sterile 20)/SPS-1 related proline/alanine-rich kinase (SPAK) and oxidative stress-response kinase-1 (OSR1) activate the renal cation cotransporters Na(+) -K(+) -2Cl(-) cotransporter (NKCC2) and Na(+) -Cl(-) cotransporter (NCC) via phosphorylation. Knockout mouse models suggest that OSR1 mainly activates NKCC2, while SPAK mainly activates NCC, with possible cross-compensation. We tested the hypothesis that disrupting both kinases causes severe polyuria and salt-wasting by generating SPAK/OSR1 double knockout (DKO) mice. DKO mice displayed lower systolic blood pressure compared with SPAK knockout (SPAK-KO) mice, but displayed no severe phenotype even after dietary salt restriction. Phosphorylation of NKCC2 at SPAK/OSR1-dependent sites was lower than in SPAK-KO mice, but still significantly greater than in wild type mice. In the renal medulla, there was significant phosphorylation of NKCC2 at SPAK/OSR1-dependent sites despite a complete absence of SPAK and OSR1, suggesting the existence of an alternative activating kinase. The distal convoluted tubule has been proposed to sense plasma [K(+) ], with NCC activation serving as the primary effector pathway that modulates K(+) secretion, by metering sodium delivery to the collecting duct. Abundance of phosphorylated NCC (pNCC) is dramatically lower in SPAK-KO mice than in wild type mice, and the additional disruption of OSR1 further reduced pNCC. SPAK-KO and kidney-specific OSR1 single knockout mice maintained plasma [K(+) ] following dietary potassium restriction, but DKO mice developed severe hypokalaemia. Unlike mice lacking SPAK or OSR1 alone, DKO mice displayed an inability to phosphorylate NCC under these conditions. These data suggest that SPAK and OSR1 are essential components of the effector pathway that maintains plasma [K(+) ].
authors.affiliation: Division of Nephrology & Hypertension, Department of Medicine, Oregon Health & Science University, Portland, OR, 97239, USA.
authors.forename: Mohammed Z
authors.initials: MZ
authors.lastname: Ferdaus
authors.affiliation: Department of Cellular & Molecular Physiology, Yale University, New Haven, CT, 06520, USA.|Systems Biology Institute, Yale University, Orange, CT, 06477, USA.
authors.forename: Karl W
authors.initials: KW
authors.lastname: Barber
authors.affiliation: INSERM U970, Paris Cardiovascular Research Center, Université Paris-Descartes, Paris, France.
authors.forename: Karen I
authors.identifier: 0000-0002-9745-0774
authors.initials: KI
authors.lastname: López-Cayuqueo
authors.affiliation: Division of Nephrology & Hypertension, Department of Medicine, Oregon Health & Science University, Portland, OR, 97239, USA.
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Terker
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México and Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico.
authors.forename: Eduardo R
authors.initials: ER
authors.lastname: Argaiz
authors.affiliation: Department of Cellular & Molecular Physiology, Yale University, New Haven, CT, 06520, USA.|Systems Biology Institute, Yale University, Orange, CT, 06477, USA.
authors.forename: Brandon M
authors.initials: BM
authors.lastname: Gassaway
authors.affiliation: INSERM U970, Paris Cardiovascular Research Center, Université Paris-Descartes, Paris, France.
authors.forename: Régine
authors.initials: R
authors.lastname: Chambrey
authors.affiliation: Molecular Physiology Unit, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México and Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, Mexico.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Gamba
authors.affiliation: Department of Cellular & Molecular Physiology, Yale University, New Haven, CT, 06520, USA.|Systems Biology Institute, Yale University, Orange, CT, 06477, USA.
authors.forename: Jesse
authors.initials: J
authors.lastname: Rinehart
authors.affiliation: Division of Nephrology & Hypertension, Department of Medicine, Oregon Health & Science University, Portland, OR, 97239, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: McCormick
mesh_terms: D000818:Animals; D001794:Blood Pressure; D006706:Homeostasis; D007686:Kidney Tubules, Distal / Q000378:metabolism* / Q000502:physiology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D011188:Potassium / Q000378:metabolism* / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D064466:Solute Carrier Family 12, Member 1 / Q000378:metabolism; D064486:Solute Carrier Family 12, Member 3 / Q000378:metabolism
chemical_list: C578715:Slc12a1 protein, mouse; C578875:Slc12a3 protein, mouse; D064466:Solute Carrier Family 12, Member 1; D064486:Solute Carrier Family 12, Member 3; C486407:Stk39 protein, mouse; C534421:OXSR1 protein, mouse; D017346:Protein Serine-Threonine Kinases; D011188:Potassium