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PKA-independent vasopressin signaling in renal collecting duct

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PKA-independent vasopressin signaling in renal collecting duct. / Datta, Arnab; Yang, Chin-Rang; Limbutara, Kavee; Chou, Chung-Lin; Rinschen, Markus M; Raghuram, Viswanathan; Knepper, Mark A.

In: FASEB J, Vol. 34, No. 5, 05.2020, p. 6129-6146.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Datta, A, Yang, C-R, Limbutara, K, Chou, C-L, Rinschen, MM, Raghuram, V & Knepper, MA 2020, 'PKA-independent vasopressin signaling in renal collecting duct', FASEB J, vol. 34, no. 5, pp. 6129-6146. https://doi.org/10.1096/fj.201902982R

APA

Datta, A., Yang, C-R., Limbutara, K., Chou, C-L., Rinschen, M. M., Raghuram, V., & Knepper, M. A. (2020). PKA-independent vasopressin signaling in renal collecting duct. FASEB J, 34(5), 6129-6146. https://doi.org/10.1096/fj.201902982R

Vancouver

Datta A, Yang C-R, Limbutara K, Chou C-L, Rinschen MM, Raghuram V et al. PKA-independent vasopressin signaling in renal collecting duct. FASEB J. 2020 May;34(5):6129-6146. https://doi.org/10.1096/fj.201902982R

Bibtex

@article{3b6bdb6c369e4ea9b4a0f8ba6837506e,
title = "PKA-independent vasopressin signaling in renal collecting duct",
abstract = "Vasopressin regulates renal water excretion by binding to a Gα s-coupled receptor (V2R) in collecting duct cells, resulting in increased water permeability through regulation of the aquaporin-2 (AQP2) water channel. This action is widely accepted to be associated with cAMP-mediated activation of protein kinase A (PKA). Here, we use phosphoproteomics in collecting duct cells in which PKA has been deleted (CRISPR-Cas9) to identify PKA-independent responses to vasopressin. The results show that V2R-mediated vasopressin signaling is predominantly, but not entirely, PKA-dependent. Upregulated sites in PKA-null cells include Ser256 of AQP2, which is critical to regulation of AQP2 trafficking. In addition, phosphorylation changes in the protein kinases Stk39 (SPAK) and Prkci (an atypical PKC) are consistent with PKA-independent regulation of these protein kinases. Target motif analysis of the phosphopeptides increased in PKA-null cells indicates that vasopressin activates one or more members of the AMPK/SNF1-subfamily of basophilic protein kinases. In vitro phosphorylation assays using recombinant, purified SNF1-subfamily kinases confirmed postulated target specificities. Of interest, measured IBMX-dependent cAMP levels were an order of magnitude higher in PKA-null than in PKA-intact cells, indicative of a PKA-dependent feedback mechanism. Overall, the findings support the conclusion that V2-receptor mediated signaling in collecting duct cells is in part PKA-independent.",
keywords = "Animals, Aquaporin 2/metabolism, Cyclic AMP-Dependent Protein Kinases/metabolism, Kidney Tubules, Collecting/cytology, Mice, Phosphoproteins/metabolism, Phosphorylation, Proteome/analysis, Receptors, Vasopressin/metabolism",
author = "Arnab Datta and Chin-Rang Yang and Kavee Limbutara and Chung-Lin Chou and Rinschen, {Markus M} and Viswanathan Raghuram and Knepper, {Mark A}",
note = "Published 2020. This article is a U.S. Government work and is in the public domain in the USA.",
year = "2020",
month = may,
doi = "10.1096/fj.201902982R",
language = "English",
volume = "34",
pages = "6129--6146",
journal = "FASEB J",
issn = "0892-6638",
publisher = "FASEB",
number = "5",

}

RIS

TY - JOUR

T1 - PKA-independent vasopressin signaling in renal collecting duct

AU - Datta, Arnab

AU - Yang, Chin-Rang

AU - Limbutara, Kavee

AU - Chou, Chung-Lin

AU - Rinschen, Markus M

AU - Raghuram, Viswanathan

AU - Knepper, Mark A

N1 - Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

PY - 2020/5

Y1 - 2020/5

N2 - Vasopressin regulates renal water excretion by binding to a Gα s-coupled receptor (V2R) in collecting duct cells, resulting in increased water permeability through regulation of the aquaporin-2 (AQP2) water channel. This action is widely accepted to be associated with cAMP-mediated activation of protein kinase A (PKA). Here, we use phosphoproteomics in collecting duct cells in which PKA has been deleted (CRISPR-Cas9) to identify PKA-independent responses to vasopressin. The results show that V2R-mediated vasopressin signaling is predominantly, but not entirely, PKA-dependent. Upregulated sites in PKA-null cells include Ser256 of AQP2, which is critical to regulation of AQP2 trafficking. In addition, phosphorylation changes in the protein kinases Stk39 (SPAK) and Prkci (an atypical PKC) are consistent with PKA-independent regulation of these protein kinases. Target motif analysis of the phosphopeptides increased in PKA-null cells indicates that vasopressin activates one or more members of the AMPK/SNF1-subfamily of basophilic protein kinases. In vitro phosphorylation assays using recombinant, purified SNF1-subfamily kinases confirmed postulated target specificities. Of interest, measured IBMX-dependent cAMP levels were an order of magnitude higher in PKA-null than in PKA-intact cells, indicative of a PKA-dependent feedback mechanism. Overall, the findings support the conclusion that V2-receptor mediated signaling in collecting duct cells is in part PKA-independent.

AB - Vasopressin regulates renal water excretion by binding to a Gα s-coupled receptor (V2R) in collecting duct cells, resulting in increased water permeability through regulation of the aquaporin-2 (AQP2) water channel. This action is widely accepted to be associated with cAMP-mediated activation of protein kinase A (PKA). Here, we use phosphoproteomics in collecting duct cells in which PKA has been deleted (CRISPR-Cas9) to identify PKA-independent responses to vasopressin. The results show that V2R-mediated vasopressin signaling is predominantly, but not entirely, PKA-dependent. Upregulated sites in PKA-null cells include Ser256 of AQP2, which is critical to regulation of AQP2 trafficking. In addition, phosphorylation changes in the protein kinases Stk39 (SPAK) and Prkci (an atypical PKC) are consistent with PKA-independent regulation of these protein kinases. Target motif analysis of the phosphopeptides increased in PKA-null cells indicates that vasopressin activates one or more members of the AMPK/SNF1-subfamily of basophilic protein kinases. In vitro phosphorylation assays using recombinant, purified SNF1-subfamily kinases confirmed postulated target specificities. Of interest, measured IBMX-dependent cAMP levels were an order of magnitude higher in PKA-null than in PKA-intact cells, indicative of a PKA-dependent feedback mechanism. Overall, the findings support the conclusion that V2-receptor mediated signaling in collecting duct cells is in part PKA-independent.

KW - Animals

KW - Aquaporin 2/metabolism

KW - Cyclic AMP-Dependent Protein Kinases/metabolism

KW - Kidney Tubules, Collecting/cytology

KW - Mice

KW - Phosphoproteins/metabolism

KW - Phosphorylation

KW - Proteome/analysis

KW - Receptors, Vasopressin/metabolism

U2 - 10.1096/fj.201902982R

DO - 10.1096/fj.201902982R

M3 - SCORING: Journal article

C2 - 32219907

VL - 34

SP - 6129

EP - 6146

JO - FASEB J

JF - FASEB J

SN - 0892-6638

IS - 5

ER -