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.
Mesh Terms:
Animals, CHO Cells, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Calcium, Cell Membrane, Cloning, Molecular, Cricetinae, Cyclic AMP, Dose-Response Relationship, Drug, GTP-Binding Proteins, Guanosine 5'-O-(3-Thiotriphosphate), Humans, Ligands, Neuropeptide Y, Oligopeptides, Peptides, Pertussis Toxin, Phenylalanine, Plasmids, Protein Binding, Protein Isoforms, Receptors, Neuropeptide Y, Signal Transduction, Temperature, Transfection, Valine
Animals, CHO Cells, Caenorhabditis elegans, Caenorhabditis elegans Proteins, Calcium, Cell Membrane, Cloning, Molecular, Cricetinae, Cyclic AMP, Dose-Response Relationship, Drug, GTP-Binding Proteins, Guanosine 5'-O-(3-Thiotriphosphate), Humans, Ligands, Neuropeptide Y, Oligopeptides, Peptides, Pertussis Toxin, Phenylalanine, Plasmids, Protein Binding, Protein Isoforms, Receptors, Neuropeptide Y, Signal Transduction, Temperature, Transfection, Valine
J Biol Chem
Date: Sep. 05, 2003
PubMed ID: 12821653
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