The proximal portion of the COOH terminus of the oxytocin receptor is required for coupling to Gq, but not Gi -: Independent mechanisms for elevating intracellular calcium concentrations from intracellular stores

As the oxytocin receptor plays a key role in parturition and lactation, there is considerable interest in defining its structure/functional relationships, We previously showed that the rat oxytocin receptor transfected into Chinese hamster ovary cells was coupled to both G(q/11) and G(i/o), and that oxytocin stimulated ERK-2 phosphorylation and prostaglandin E-2 synthesis via protein kinase C activity. In this study, we show that deletion of 51 amino acid residues from the carboxyl terminus resulted in reduced affinity for oxytocin and a corresponding rightward shift in the dose-response curve for oxytocin-stimulated [Ca2+](i). However, oxytocin-stimulated ERK-2 phosphorylation and prostaglandin E-2 synthesis did not occur in cells expressing the truncated receptor. Oxytocin also failed to increase phospholipase A activity or activate protein kinase C, indicating that the mutant receptor is uncoupled from G(q)-mediated pathways. The Delta 51 receptor is coupled to G(i), as oxytocin-stimulated Ca2+ transients were inhibited by pertussis toxin, and a G beta gamma sequestrant. Preincubation of Delta 51 cells with the tyrosine kinase inhibitor, genistein, also blocked the oxytocin effect. A Delta 39 mutant had all the activities of the wild type oxytocin receptor, These results show that the portion between 39 and 51 residues from the COOH terminus of the rat oxytocin receptor is required for interaction with G(q/11), but not G(i/o). Furthermore, an increase in intracellular calcium was generated via a G(i)beta gamma-tyrosine kinase pathway from intracellular stores that are distinct from G(q)-mediated inositol trisphosphate-regulated stores.