A clone PKSV-PCT Cl.10 referred to as Cl.10 was selected from the PKSV-PCT renal proximal tubule cell line which expressed peptide YY (PYY) receptors (Voisin, T., Bens, M., Cluzeaud, F., Vandewalle, A., and Laburthe, M.(1993) J. Biol. Chem. 268, 20547-20554). In order to identify G protein(s) coupled to PYY receptors, antisense G protein RNAs were expressed in Cl.10 cells by transfecting the pcDNA3 vector into which were inserted 39 bases of the 5`-noncoding region of G or G used as specific antisense templates. A Cl.10/ clone was selected which displayed a drastic decrease (>90%) of the expression of G without changes of G, G, and G subunits (G is not present in Cl.10 cells) as evidenced by Western blots. When compared to untransfected cells, this clone exhibited: (i) an increase in the dissociation constant of PYY receptors (5.3 versus 0.6 nM) identical to that observed in pertussis toxin-treated untransfected cells; (ii) an absence of inhibition of I-PYY binding by guanosine 5`-O-(thiotriphosphate) (GTPS); and (iii) the failure of PYY to inhibit cAMP levels and to stimulate [methyl-H]thymidine incorporation into DNA. A clone was also selected which exhibited a specific decrease (>80%) of G as compared to untransfected cells. The sensitivity to GTPS and the dissociation constant of PYY receptors as well as PYY-mediated inhibition of cAMP were identical to those observed in untransfected cells. These findings support an exclusive coupling of PYY receptors to G.

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