title: Isolation and genetic characterization of a renal epithelial cell mutant defective in vasopressin (V2) receptor binding and function.
abstract: A novel mutant of the LLC-PK1 renal epithelial cell line, VPR1, was isolated after mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine and selection using a photoactivatable vasopressin analogue [1-(3-mercapto)propionic acid, 8-(N6-4-azidophenylamidino)lysine] vasopressin. The VPR1 mutant cell line possessed less than 5% parental V2 receptor binding for vasopressin but exhibited normal calcitonin receptor binding. In contrast to LLC-PK1 cells (wild type), VPR1 cells exhibited no response to vasopressin in terms of in vitro adenylate cyclase activation, in vivo cAMP production, or urokinase-type plasminogen activator induction. The responses of VPR1 cells to other agents, such as calcitonin, the adenylate cyclase activator forskolin, the GTP analogue guanosine 5'-[beta, gamma-imino] triphosphate, 8-bromo adenosine-3',5'-monophosphate were comparable to those of the parental cell line. Somatic cell hybrids were derived from the cell lines LLC-PK1 and VPR1 and analyzed for the dominance/recessiveness of the VPR1 mutant phenotype. Hybrids were found to possess normal vasopressin binding activity as well as functional responses to the hormone, indicating that the mutation affecting the V2 receptor in VPR1 cells is recessive. The VPR1 cell line may thus have application as a recipient for the expression of the V2 receptor gene using DNA-transfer.
authors.affiliation: Max-Planck-Institut für Biophysik, Frankfurt am Main, Federal Republic of Germany.
authors.forename: H
authors.initials: H
authors.lastname: Luzius
authors.forename: D A
authors.initials: DA
authors.lastname: Jans
authors.forename: P
authors.initials: P
authors.lastname: Jans
authors.forename: F
authors.initials: F
authors.lastname: Fahrenholz
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D005343:Fibrinolytic Agents / Q000378:metabolism; D006822:Hybrid Cells; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D009154:Mutation; D010960:Plasminogen Activators / Q000378:metabolism; D011945:Receptors, Angiotensin / Q000235:genetics* / Q000378:metabolism; D017483:Receptors, Vasopressin; D013552:Swine; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism; D014667:Vasopressins / Q000378:metabolism*
chemical_list: D005343:Fibrinolytic Agents; D011945:Receptors, Angiotensin; D017483:Receptors, Vasopressin; D014667:Vasopressins; D008769:Methylnitronitrosoguanidine; D000242:Cyclic AMP; D010960:Plasminogen Activators; D014568:Urokinase-Type Plasminogen Activator; D000262:Adenylyl Cyclases