Activation of phosphatidylinositol 4,5-bisphosphate supply by agonists and non-hydrolysable GTP analogues

Abstract

PtdIns(4,5)P2 serves as a precursor of a diverse family of signalling molecules, including diacylglycerol (and hence phosphatidic acid), Ins(1,4,5)P3 [and hence Ins(1,3,4,5)P4] and PtdIns(3,4,5)P3. The production of these messengers can be activated by agonists, and therefore the rate of utilization of PtdIns(4,5)P2 can vary dramatically. Although cells can only meet these large changes in demand for PtdIns(4,5)P2 by increasing its synthesis and/or by continuously cycling it at a rate that exceeds its potential consumption (avoiding the need for a co-ordinated activation mechanism), no satisfactory explanation for how this is achieved in agonist-stimulated cells has yet been provided. We show here that, in streptolysin-O-permeabilized neutrophils, N-formylmethionyl-leucyl-phenylalanine (FMLP), platelet-activating factor (PAF) and non-hydrolysable GTP analogues can cause large activations of PtdIns4P 5-kinase, suggesting that cells can accommodate agonist-activated rates of consumption of PtdIns(4,5)P2 without having to sustain continuous, comparably rapid and energetically expensive ‘futile cycling’ reactions.