A protein kinase A-independent pathway controlling aquaporin 2 trafficking as a possible cause for the syndrome of inappropriate antidiuresis associated with polycystic kidney disease 1 haploinsufficiency.
Renal water reabsorption is controlled by arginine vasopressin (AVP), which binds to V2 receptors, resulting in protein kinase A (PKA) activation, phosphorylation of aquaporin 2 (AQP2) at serine 256, and translocation of AQP2 to the plasma membrane. However, AVP also causes dephosphorylation of AQP2 at S261. Recent studies showed that cyclin-dependent kinases (cdks) can phosphorylate AQP2 peptides at S261 in vitro. We investigated the possible role of cdks in the phosphorylation of AQP2 and identified a new PKA-independent pathway regulating AQP2 trafficking. In ex vivo kidney slices and MDCK-AQP2 cells, R-roscovitine, a specific inhibitor of cdks, increased pS256 levels and decreased pS261 levels. The changes in AQP2 phosphorylation status were paralleled by increases in cell surface expression of AQP2 and osmotic water permeability in the absence of forskolin stimulation. R-Roscovitine did not alter cAMP-dependent PKA activity but specifically reduced protein phosphatase 2A (PP2A) expression and activity in MDCK cells. Notably, we found reduced PP2A expression and activity and reduced pS261 levels in Pkd1(+/-) mice displaying a syndrome of inappropriate antidiuresis with high levels of pS256, despite unchanged AVP and cAMP. Similar to previous findings in Pkd1(+/-) mice, R-roscovitine treatment caused a significant decrease in intracellular calcium in MDCK cells. Our data indicate that reduced activity of PP2A, secondary to reduced intracellular Ca(2+) levels, promotes AQP2 trafficking independent of the AVP-PKA axis. This pathway may be relevant for explaining pathologic states characterized by inappropriate AVP secretion and positive water balance.
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy; grazia.tamma@uniba.it.
Grazia
G
Tamma
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy;
Domenica
D
Lasorsa
Department of Physiology, Radboud University Medical Centre, Nijmegen, The Netherlands; and.
Christiane
C
Trimpert
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy;
Marianna
M
Ranieri
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy;
Annarita
A
Di Mise
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy;
Maria Grazia
MG
Mola
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy;
Lisa
L
Mastrofrancesco
Institute of Physiology, University of Zurich, Zurich, Switzerland.
Olivier
O
Devuyst
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy;
Maria
M
Svelto
Department of Physiology, Radboud University Medical Centre, Nijmegen, The Netherlands; and.
Peter M T
PM
Deen
Department of Biosciences Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy;
Giovanna
G
Valenti
D000818:Animals; D051399:Aquaporin 2 / Q000378:metabolism*; D020013:Calcium Signaling; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D057895:Haploinsufficiency; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D007690:Polycystic Kidney Diseases / Q000201:enzymology* / Q000235:genetics; D054648:Protein Phosphatase 2 / Q000378:metabolism*; D011687:Purines; D017207:Rats, Sprague-Dawley; D000077546:Roscovitine; D014667:Vasopressins / Q000378:metabolism
C496025:Aqp2 protein, rat; D051399:Aquaporin 2; D011687:Purines; D000077546:Roscovitine; D014667:Vasopressins; D017868:Cyclic AMP-Dependent Protein Kinases; D018844:Cyclin-Dependent Kinases; D054648:Protein Phosphatase 2