Molecular Cloning and Expression of an Avian G Protein-Coupled P2Y Receptor
- Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, North Carolina 27599
Abstract
A family of G protein-coupled P2Y receptors that are activated by adenine and uridine nucleotides has been identified recently. Degenerate primers based on conserved sequences in these P2Y receptors were used to amplify turkey DNA, which was used to isolate the complete coding sequence of a cDNA that encodes a novel G protein-coupled receptor. Stable expression of this avian cDNA in 1321N1 human astrocytoma cells resulted in the conveyance of marked inositol phosphate responses to various nucleotides. Although this cloned avian receptor exhibited its highest homology to the previously cloned mammalian P2Y4 receptor, its pharmacological selectivity was not consistent with the avian receptor’s being a species homologue of the P2Y4 receptor. That is, whereas the P2Y4receptor is selectively activated by UTP and is not activated by ATP or AP4A, the novel avian receptor was potently activated by ATP and AP4A as well as by UTP. Taken together, these results describe the identification of an avian phospholipase C-coupled P2Y receptor that, like the mammalian P2Y2 receptor, is activated by both adenine and uridine nucleotides.
Footnotes
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Send reprint requests to: José L. Boyer, Department of Pharmacology, CB# 7365, Mary Ellen Jones Bldg., University of North Carolina, Chapel Hill, NC 27599. E-mail:boyerl{at}med.unc.edu
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↵3 F. del Toro, J. L. Boyer, T. K. Harden, and R. A. Nicholas, manuscript in preparation.
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This work was supported by United States Public Health Service Grants HL54889 and GM38213.
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↵1 The nomenclature used here is that suggested by the International Union of Pharmacology Committee on Receptor Nomenclature and Drug Classification (32) and adapted by the Purinoceptor Subcommittee of the same organization (33).
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↵2 Q. Li, M. Olesky, T. K. Harden, and R. A. Nicholas, manuscript in preperation.
- Abbreviations:
- 2MeSATP
- 2-methylthio-ATP
- 2MeSADP
- 2-methylthio-ADP
- EGTA
- ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid
- DMEM
- Dulbecco’s modified Eagle’s medium
- HEPES
- 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- PIPES
- piperazine-N,N′-bis-2-ethanesulfonic acid
- APxA
- diadenosine x-phosphate, wherex is the number of phosphates
- PCR
- polymerase chain reaction
- bp
- base pair(s)
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- Received July 9, 1997.
- Accepted August 25, 1997.
- The American Society for Pharmacology and Experimental Therapeutics
