title: A Systems Level Analysis of Vasopressin-mediated Signaling Networks in Kidney Distal Convoluted Tubule Cells.
abstract: The kidney distal convoluted tubule (DCT) plays an essential role in maintaining body sodium balance and blood pressure. The major sodium reabsorption pathway in the DCT is the thiazide-sensitive NaCl cotransporter (NCC), whose functions can be modulated by the hormone vasopressin (VP) acting via uncharacterized signaling cascades. Here we use a systems biology approach centered on stable isotope labeling by amino acids in cell culture (SILAC) based quantitative phosphoproteomics of cultured mouse DCT cells to map global changes in protein phosphorylation upon acute treatment with a VP type II receptor agonist 1-desamino-8-D-arginine vasopressin (dDAVP). 6330 unique proteins, containing 12333 different phosphorylation sites were identified. 185 sites were altered in abundance following dDAVP. Basophilic motifs were preferential targets for upregulated sites upon dDAVP stimulation, whereas proline-directed motifs were prominent for downregulated sites. Kinase prediction indicated that dDAVP increased AGC and CAMK kinase families' activities and decreased activity of CDK and MAPK families. Network analysis implicated phosphatidylinositol-4,5-bisphosphate 3-kinase or CAMKK dependent pathways in VP-mediated signaling; pharmacological inhibition of which significantly reduced dDAVP induced increases in phosphorylated NCC at an activating site. In conclusion, this study identifies unique VP signaling cascades in DCT cells that may be important for regulating blood pressure.
authors.affiliation: Center for Interactions of Proteins in Epithelial Transport, Department of Biomedicine, Aarhus University, 8000, Aarhus, Denmark.
authors.forename: Lei
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Center for Interactions of Proteins in Epithelial Transport, Department of Biomedicine, Aarhus University, 8000, Aarhus, Denmark.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wu
authors.affiliation: Center for Interactions of Proteins in Epithelial Transport, Department of Biomedicine, Aarhus University, 8000, Aarhus, Denmark.
authors.forename: Marleen L A
authors.initials: MLA
authors.lastname: Kortenoeven
authors.affiliation: Center for Interactions of Proteins in Epithelial Transport, Department of Biomedicine, Aarhus University, 8000, Aarhus, Denmark.|Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Trairak
authors.initials: T
authors.lastname: Pisitkun
authors.affiliation: Center for Interactions of Proteins in Epithelial Transport, Department of Biomedicine, Aarhus University, 8000, Aarhus, Denmark.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Fenton
mesh_terms: D000818:Animals; D001794:Blood Pressure / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D003894:Deamino Arginine Vasopressin / Q000494:pharmacology; D053503:Epithelial Sodium Channels / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D017136:Ion Transport / Q000187:drug effects; D007553:Isotope Labeling; D007686:Kidney Tubules, Distal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation / Q000187:drug effects; D017483:Receptors, Vasopressin / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D012964:Sodium / Q000378:metabolism; D064486:Solute Carrier Family 12, Member 3 / Q000235:genetics / Q000378:metabolism; D049490:Systems Biology; D049971:Thiazides / Q000494:pharmacology; D046509:Tissue Culture Techniques; D014667:Vasopressins / Q000494:pharmacology*
chemical_list: D053503:Epithelial Sodium Channels; D017483:Receptors, Vasopressin; C507047:Scnn1a protein, mouse; C507049:Scnn1b protein, mouse; C578875:Slc12a3 protein, mouse; D064486:Solute Carrier Family 12, Member 3; D049971:Thiazides; D014667:Vasopressins; D012964:Sodium; D054737:Calcium-Calmodulin-Dependent Protein Kinase Kinase; D003894:Deamino Arginine Vasopressin; D002118:Calcium