Role of the human V1 vasopressin receptor COOH terminus in internalization and mitogenic signal transduction.
We studied the role played by the intracellular COOH-terminal region of the human arginine vasopressin (AVP) V1-vascular receptor (V1R) in ligand binding, trafficking, and mitogenic signal transduction in Chinese hamster ovary cells stably transfected with the human AVP receptor cDNA clones that we had isolated previously. Truncations, mutations, or chimeric alterations of the V1R COOH terminus did not alter ligand binding, but agonist-induced V1R internalization and recycling were reduced in the absence of the proximal region of the V(1)R COOH terminus. Coupling to phospholipase C was altered as a function of the COOH-terminal length. Deletion of the proximal portion of the V1R COOH terminus or its replacement by the V2-renal receptor COOH terminus prevented AVP stimulation of DNA synthesis and progression through the cell cycle. Mutation of a kinase consensus motif in the proximal region of the V1R COOH terminus also abolished the mitogenic response. Thus the V1R cytoplasmic COOH terminus is not involved in ligand specificity but is instrumental in receptor trafficking and facilitates the interaction between the intracellular loops of the receptor, G protein, and phospholipase C. It is absolutely required for transmission of the mitogenic action of AVP, probably via a specific kinase phosphorylation site.
Thibonnier
M
M
Division of Clinical and Molecular Endocrinology, Department of Medicine, University Hospitals of Cleveland and Case Western Reserve University School of Medicine, Cleveland, Ohio, 44106, USA. mxt10@po.cwru.edu
Plesnicher
C L
CL
Berrada
K
K
Berti-Mattera
L
L
D000595:Amino Acid Sequence; D000818:Animals; D001127:Arginine Vasopressin / Q000378:metabolism; D001667:Binding, Competitive / Q000187:drug effects; D016466:CHO Cells; D002264:Carboxylic Acids / Q000737:chemistry*; D002453:Cell Cycle / Q000502:physiology; D006224:Cricetinae; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D005434:Flow Cytometry; D006801:Humans; D007202:Indicators and Reagents; D007295:Inositol Phosphates / Q000096:biosynthesis; D008938:Mitosis / Q000502:physiology*; D008969:Molecular Sequence Data; D011869:Radioligand Assay; D017483:Receptors, Vasopressin / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*; D013936:Thymidine / Q000378:metabolism
D002264:Carboxylic Acids; D007202:Indicators and Reagents; D007295:Inositol Phosphates; D017483:Receptors, Vasopressin; D001127:Arginine Vasopressin; D004247:DNA; D013936:Thymidine