CLONING AND TISSUE DISTRIBUTION OF THE HUMAN P2Y(1) RECEPTOR

Sets of degenerate oligonucleotide primers synthesized on the basis of the best conserved regions of the chick brain P-2Y/P2Y(1) and the mur ine neuroblastoma P-2U/P2Y(2) receptors were used in polymerase chain reaction experiments on human genomic DNA. An amplified fragment of 71 2 base pairs was then used as a probe to screen a human genomic DNA li brary. Several clones were isolated and sequencing revealed an intronl ess 1122 base pair open reading frame. The corresponding amino acid se quence revealed 83% identity with the chick brain P2Y(1) receptor and 34% with the murine neuroblastoma P2Y(2) receptor. In COS-7 cells tran sfected with the coding sequence inserted into the pcDNA3 expression v ector, 2-methylthioATP and ATP produced a strong stimulation of inosit ol phosphates, a typical response of a P2Y(1) receptor. Northern blot analysis detected a 6.7 kilobase messenger RNA in most human tissues, the strongest signals being observed in prostate and ovary. (C) 1996 A cademic Press, Inc.