Cloning and tissue distribution of the human P2Y1 receptor.
Sets of degenerate oligonucleotide primers synthesized on the basis of the best conserved regions of the chick brain P2Y/P2Y1 and the murine neuroblastoma P2U/P2Y2 receptors were used in polymerase chain reaction experiments on human genomic DNA. An amplified fragment of 712 base pairs was then used as a probe to screen a human genomic DNA library. Several clones were isolated and sequencing revealed an intronless 1122 base pair open reading frame. The corresponding amino acid sequence revealed 83% identity with the chick brain P2Y1 receptor and 34% with the murine neuroblastoma P2Y2 receptor. In COS-7 cells transfected with the coding sequence inserted into the pcDNA3 expression vector, 2-methylthioATP and ATP produced a strong stimulation of inositol phosphates, a typical response of a P2Y1 receptor. Northern blot analysis detected a 6.7 kilobase messenger RNA in most human tissues, the strongest signals being observed in prostate and ovary.
Janssens
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Institute of Interdisciplinary Research, School of Medicine, Erasme Hospital, Université Libre de Bruxelles, Belgium. rajansse@ulb.ac.be.
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Pirotton
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Samson
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Parmentier
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Boeynaems
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular; D004247:DNA; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D018048:Receptors, Purinergic P2 / Q000235:genetics* / Q000378:metabolism; D058472:Receptors, Purinergic P2Y1; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
C546382:P2RY1 protein, human; C546383:P2ry1 protein, mouse; D018048:Receptors, Purinergic P2; D058472:Receptors, Purinergic P2Y1; D004247:DNA