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J Biol Chem, Vol. 273, Issue 21, 12710-12715, May 22, 1998

Phosphatidic Acid-induced Elevation of Intracellular Ca2+ Is Mediated by RhoA and H2O2 in Rat-2 Fibroblasts

Zee-Won LeeDagger , Soo-Mi KweonDagger , Byung-Chul Kim§, Sun-Hee Leem, Incheol Shinparallel , Jae-Hong Kim§, and Kwon-Soo HaDagger

From the Dagger  Biomolecule Research Group, Korea Basic Science Institute, Taejon 305-333, Korea, the § Laboratory of Molecular and Cellular Genetics, Institute of Environment and Life Science, Hallym University, Chun-Cheon, Kangwon-do 200-702, Korea, the  Department of Biology, Dong-A University, Pusan 604-714, Korea, and the parallel  Department of Biochemistry, College of Medicine, Hanyang University, Seoul 133-701, Korea

We have investigated possible roles of RhoA and H2O2 in the elevation of intracellular Ca2+ ([Ca2+]i) by phosphatidic acid (PA) in Rat-2 fibroblasts. PA induced a transient elevation of [Ca2+]i in the presence or absence of EGTA. Lysophosphatidic acid (LPA) also increased [Ca2+]i, but the sustained Ca2+ response was inhibited by EGTA. LPA stimulated the production of inositol phosphates, but PA did not. In the presence of EGTA, preincubation with LPA completely blocked the subsequent elevation of [Ca2+]i by PA, but not vice versa. PA stimulated the translocation of RhoA to the particulate fraction as did LPA. Scrape loading of C3 transferase inhibited the transient Ca2+ response to PA, but not to LPA, suggesting an essential role of RhoA in the elevation of [Ca2+]i by PA. H2O2 also induced a transient increase of [Ca2+]i as did PA. H2O2 scavengers, catalase and N-acetyl-L-cysteine, completely blocked the rise of [Ca2+]i stimulated by PA, but not by LPA. Furthermore, preincubation with PA blocked the subsequent Ca2+ response to H2O2, and the incubation with H2O2 also blocked the PA-induced rise of [Ca2+]i. Thus, it was suggested that PA stimulated Ca2+ release from PA-sensitive, but not inositol 1,4,5-trisphosphate-sensitive, Ca2+ stores by the activation of RhoA and intracellular H2O2.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.


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