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Arginine Vasopressin regulates the renal Na-Cl and Na-K-Cl Cotransporters through With-No-Lysine Kinase 4 and Inhibitor 1 Phosphorylation


Carbajal-Contreras, Héctor; Murillo-de-Ozores, Adrian Rafael; Magaña-Avila, Germán; Marquez-Salinas, Alejandro; Bourqui, Laurent; Tellez-Sutterlin, Michelle; Bahena-Lopez, Jessica P; Cortes-Arroyo, Eduardo; González-Behn-Eschemburg, Sebastián; Lopez-Saavedra, Alejandro; Vazquez, Norma; Ellison, David H; Loffing, Johannes; Gamba, Gerardo; Castañeda-Bueno, María (2024). Arginine Vasopressin regulates the renal Na-Cl and Na-K-Cl Cotransporters through With-No-Lysine Kinase 4 and Inhibitor 1 Phosphorylation. American Journal of Physiology : Renal Physiology, 326(2):F285-F299.

Abstract

Vasopressin regulates water homeostasis via the V2 receptor in the kidney at least in part through protein kinase A (PKA) activation. Vasopressin, through an unknown pathway, upregulates the activity and phosphorylation of the Na+-Cl- cotransporter (NCC) and Na+-K+-2Cl- cotransporter 2 (NKCC2) by Ste20-related Proline/Alanine rich Kinase (SPAK) and Oxidative Stress Responsive kinase 1 (OSR1), which are regulated by the With No Lysine (K) kinase (WNK) family. Phosphorylation of WNK4 at PKA consensus motifs may be involved. Inhibitor 1 (I1), a Protein Phosphatase 1 (PP1) inhibitor, may also play a role. In HEK293 cells, we assessed the phosphorylation of WNK4, SPAK, NCC, or NKCC2 in response to forskolin or desmopressin. WNK4 and cotransporter phosphorylation was studied in desmopressin-infused WNK4$^{-/-}$ mice and in tubule suspensions. In HEK293 cells, only wild-type WNK4, but not WNK1, WNK3, or a WNK4 mutant lacking PKA phosphorylation motifs could upregulate SPAK or cotransporter phosphorylation in response to forskolin or desmopressin. I1 transfection maximized SPAK phosphorylation in response to forskolin in the presence of WNK4, but not of mutant WNK4 lacking PP1 regulation. We observed direct PP1 regulation of NKCC2 dephosphorylation, but not of NCC or SPAK in the absence of WNK4. WNK4$^{-/-}$ mice with desmopressin treatment did not increase SPAK/OSR1, NCC, or NKCC2 phosphorylation. In stimulated tubule suspensions from WNK4$^{-/-}$ mice, upregulation of pNKCC2 was reduced, whereas upregulation of pSPAK was absent. These findings suggest that WNK4 is a central node in which kinase and phosphatase signaling converge to connect cAMP signaling to the SPAK/OSR1-NCC/NKCC2 pathway.

Abstract

Vasopressin regulates water homeostasis via the V2 receptor in the kidney at least in part through protein kinase A (PKA) activation. Vasopressin, through an unknown pathway, upregulates the activity and phosphorylation of the Na+-Cl- cotransporter (NCC) and Na+-K+-2Cl- cotransporter 2 (NKCC2) by Ste20-related Proline/Alanine rich Kinase (SPAK) and Oxidative Stress Responsive kinase 1 (OSR1), which are regulated by the With No Lysine (K) kinase (WNK) family. Phosphorylation of WNK4 at PKA consensus motifs may be involved. Inhibitor 1 (I1), a Protein Phosphatase 1 (PP1) inhibitor, may also play a role. In HEK293 cells, we assessed the phosphorylation of WNK4, SPAK, NCC, or NKCC2 in response to forskolin or desmopressin. WNK4 and cotransporter phosphorylation was studied in desmopressin-infused WNK4$^{-/-}$ mice and in tubule suspensions. In HEK293 cells, only wild-type WNK4, but not WNK1, WNK3, or a WNK4 mutant lacking PKA phosphorylation motifs could upregulate SPAK or cotransporter phosphorylation in response to forskolin or desmopressin. I1 transfection maximized SPAK phosphorylation in response to forskolin in the presence of WNK4, but not of mutant WNK4 lacking PP1 regulation. We observed direct PP1 regulation of NKCC2 dephosphorylation, but not of NCC or SPAK in the absence of WNK4. WNK4$^{-/-}$ mice with desmopressin treatment did not increase SPAK/OSR1, NCC, or NKCC2 phosphorylation. In stimulated tubule suspensions from WNK4$^{-/-}$ mice, upregulation of pNKCC2 was reduced, whereas upregulation of pSPAK was absent. These findings suggest that WNK4 is a central node in which kinase and phosphatase signaling converge to connect cAMP signaling to the SPAK/OSR1-NCC/NKCC2 pathway.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Anatomy
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Uncontrolled Keywords:Protein Kinase A; Protein Phosphatase 1; distal convoluted tubule; thick ascending limb; urinary concentration
Language:English
Date:1 February 2024
Deposited On:09 Jan 2024 11:48
Last Modified:30 Apr 2024 01:46
Publisher:American Physiological Society
ISSN:1522-1466
OA Status:Closed
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1152/ajprenal.00343.2023
PubMed ID:38096266