Differential regulation of P2Y(11) receptor-mediated signalling to phospholipase C and adenylyl cyclase by protein kinase C in HL-60 promyelocytes

Abstract

1 The regulatory mode of the P2Y(11) purinoceptor-mediated signalling cascades towards phospholipase C and adenylyl cyclase was studied in HL-60 promyelocytes. 2 Treatment with the potent P2Y(11) receptor activator dATP evoked an elevated intracellular Ca2+ concentration ([Ca2+],) and inositol 1,4,5-trisphosphate (IP3) production that was sustained for longer than 30 min. However, the dATP-induced responses were significantly inhibited by the activation of protein kinase C after a short exposure to phorbol 12-myristate 13-acetate (PMA). 3 dATP also potently stimulated cyclic AMP production with half maximum effect seen at 23 +/- 7 mu M dATP. In addition, a 5-min pretreatment with PMA enhanced the dATP-stimulated cyclic AMP accumulation. 4 PMA potentiated the cyclic AMP production when adenylyl cyclase was activated directly by forskolin or indirectly by G protein activation after cholera toxin treatment. dATP also enhanced the forskolin-mediated cyclic AMP generation. 5 Treatment of the cells with 10 mu M U-73122, which almost completely blocked the dATP-stimulated IF, production and [Ca2+](i) rise, had no effect on cyclic AMP accumulation, while 10 mu M 9-(tetrahydro-2-furyl)adenine (SQ 22536), which inhibited the adenylyl cyclase activation; did not effect the dATP-stimulated phosphoinositide turnover. 6 Taken together, the results indicate that P2Y(11) receptor-mediated activation of phospholipase C and adenylyl cyclase occurs through independent pathways and is differentially regulated by protein kinase C in HL-60 cells.