Attenuation of LPA-Mediated Calcium Signaling and Inositol Polyphosphate Production in RAT-1 Fibroblasts Transformed by the V-SRC Oncogene

Abstract

Alterations in cellular signaling underlie the transforming actions of many oncogenes. The vsrc oncogene tyrosine kinase, pp60vsrc, is known to alter multiple signal transduction pathways, including those involving phosphatidylinositol (PI) metabolism. In this study, we investigated the effects of vsrc-transformation on lysophosphatidic acid (LPA) receptor coupling to intracellular free calcium [Ca2+]iand PI turnover in rat-1 fibroblasts. In normal rat-1 cells, LPA rapidly elevated [Ca2+]i(EC50= 10nM). In contrast, the ability of LPA to mobilize calcium was markedly attenuated in rat-1-vsrccells. Further study revealed that the LPA-mediated generation of inositol (1,4,5)P3and other inositol polyphosphates was also markedly attenuated in the vsrc-transformed cells. Although LPA caused a transient reduction in the level of PI(4,5)P2in normal rat-1 cells, the agonist elevated the level of PI(4,5)P2in the vsrc-transformed cells. These findings demonstrate that vsrc-transformation alters the coupling of LPA receptors to PI turnover and calcium signaling in rat-1 cells, and point to G protein-coupled receptor systems as targets for modulation by the vsrc kinase.