Differential activation of "social" and "solitary" variants of the Caenorhabditis elegans G protein-coupled receptor NPR-1 by its cognate ligand AF9.
Natural variations of wild Caenorhabditis elegans isolates having either Phe-215 or Val-215 in NPR-1, a putative orphan neuropeptide Y-like G protein-coupled receptor, result in either "social" or "solitary" feeding behaviors (de Bono, M., and Bargmann, C. I. (1998) Cell 94, 679-689). We identified a nematode peptide, GLGPRPLRF-NH2 (AF9), as a ligand activating the cloned NPR-1 receptor heterologously expressed in mammalian cells. Shifting cell culture temperatures from 37 to 28 degrees C, implemented 24 h after transfections, was essential for detectable functional expression of NPR-1. AF9 treatments linked both cloned receptor variants to activation of Gi/Go proteins and cAMP inhibition, thus allowing for classification of NPR-1 as an inhibitory G protein-coupled receptor. The Val-215 receptor isoform displayed higher binding and functional activity than its Phe-215 counterpart. This finding parallels the in vivo observation of a more potent repression of social feeding by the npr-1 gene encoding the Val-215 form of the receptor, resulting in dispersing (solitary) animals. Since neuropeptide Y shows no sequence homology to AF9 and was functionally inactive at the cloned NPR-1, we propose to rename NPR-1 and refer to it as an AF9 receptor, AF9-R1.
Kubiak
Teresa M
TM
Animal Health Discovery Research, Pharmacia Corp., Kalamazoo, Michigan 49001, USA. teresa.m.kubiak@pfizer.com
Larsen
Martha J
MJ
Nulf
Susan C
SC
Zantello
Marjorie R
MR
Burton
Katherine J
KJ
Bowman
Jerry W
JW
Modric
Tomislav
T
Lowery
David E
DE
D000818:Animals; D016466:CHO Cells; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D019204:GTP-Binding Proteins / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D008024:Ligands; D009478:Neuropeptide Y / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D037342:Pertussis Toxin / Q000494:pharmacology; D010649:Phenylalanine / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms; D017476:Receptors, Neuropeptide Y / Q000737:chemistry* / Q000378:metabolism; D015398:Signal Transduction; D013696:Temperature; D014162:Transfection; D014633:Valine / Q000737:chemistry
C477737:AF9 peptide, C elegans; D029742:Caenorhabditis elegans Proteins; D008024:Ligands; C468272:NPR-1 protein, C elegans; D009478:Neuropeptide Y; D009842:Oligopeptides; D010455:Peptides; D020033:Protein Isoforms; D017476:Receptors, Neuropeptide Y; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D010649:Phenylalanine; D000242:Cyclic AMP; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D014633:Valine; D002118:Calcium