title: Overlapping genes in Nalp6/PYPAF5 locus encode two V2-type vasopressin isoreceptors: angiotensin-vasopressin receptor (AVR) and non-AVR.
abstract: The angiotensin-vasopressin receptor (AVR) responds with equivalent affinities to angiotensin II (ANG II) and vasopressin and is coupled to adenylate cyclase and hence a V2-type vasopressin receptor. AVR maps to the Nalp6 locus and overlaps with the larger Nalp6/PYPAF5 reported to be a T cell/granulocyte-specific, cytoplasmic-specific proapoptotic protein, thus questioning the existence of AVR. Here we confirm, through different experimental modalities, that AVR is distinct from Nalp6/PYPAF5 based on different mRNA and protein sizes, subcellular localization, and tissue-specific expression patterns. Binding studies of PYPAF5-specific Cos1 transfectants detect high-affinity binding to vasopressin but not ANG II, thus assigning PYPAF5 as a non-AVR (NAVR). Signaling array analysis reveals that AVP stimulation of AVR- and NAVR-specific Cos1 transfectants results in diametrical activation as well as coactivation of signaling pathways known to mediate renal sodium and water balance. Likewise, ANG II stimulation of Cos1-AVR transfectants reveals a signaling profile distinct from that of AVP-stimulated Cos1-AVR transfectants. Analysis of genomic organization of the AVR/NAVR locus shows an overlapping gene arrangement with alternative promoter usage resulting in different NH(2) termini for NAVR and AVR. In addition to core promoter elements, androgen and estrogen response elements are detected. Promoter analysis of NAVR/AVR 5'-regulatory region detects transcriptional upregulation by testosterone and synergistic upregulation by testosterone and estrogen, thus suggesting that AVR and/or NAVR contribute to sex-specific V2-type vasopressin-mediated effects. Altogether, confirmation of AVR and identification of NAVR as vasopressin receptors are concordant with emerging vasopressin functions not attributable to V1a, V1b, or V2 receptors and add molecular bases for the multifunctional complexity of vasopressin-mediated functions and regulation.
authors.affiliation: Section of Molecular Medicine, Department of Medicine and Whitaker Cardiovascular Institute, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Victoria L M
authors.initials: VL
authors.lastname: Herrera
authors.forename: Pia
authors.initials: P
authors.lastname: Bagamasbad
authors.forename: Tamara
authors.initials: T
authors.lastname: Didishvili
authors.forename: Julius L
authors.initials: JL
authors.lastname: Decano
authors.forename: Nelson
authors.initials: N
authors.lastname: Ruiz-Opazo
mesh_terms: D000804:Angiotensin II / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D005786:Gene Expression Regulation / Q000187:drug effects; D016678:Genome / Q000235:genetics; D012739:Gonadal Steroid Hormones / Q000494:pharmacology; D008024:Ligands; D018613:Microscopy, Confocal; D009928:Organ Specificity / Q000187:drug effects; D010455:Peptides / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011945:Receptors, Angiotensin / Q000235:genetics* / Q000378:metabolism; D017483:Receptors, Vasopressin / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D024363:Transcription Initiation Site; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014667:Vasopressins / Q000378:metabolism
chemical_list: D012739:Gonadal Steroid Hormones; D008024:Ligands; C456360:Nlrp6 protein, rat; D010455:Peptides; D020033:Protein Isoforms; D012333:RNA, Messenger; D011945:Receptors, Angiotensin; D017483:Receptors, Vasopressin; D014667:Vasopressins; D000804:Angiotensin II