Colocalization of Ras and Ral on the membrane is required for Ras-dependent Ral activation through Ral GDP dissociation stimulator

• Published in: Oncogene Vol. 15; no. 24; pp. 2899 - 2907
• Main Authors: Kishida, S, Koyama, S, Matsubara, K, Kishida, M, Matsuura, Y, Kikuchi, A
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 11.12.1997
• Subjects: Animals; Cell activation; Cell Membrane - metabolism; COS Cells; Epidermal growth factor; GTP-Binding Proteins - genetics; GTP-Binding Proteins - metabolism; GTP-Binding Proteins - physiology; Guanosine diphosphate; Inositol; Lecithin; Liposomes; Liposomes - metabolism; Phosphatidylcholine; Phosphatidylethanolamine; Phosphatidylserine; Phospholipids; Phospholipids - physiology; Post-translation; Protein Processing, Post-Translational; ral Guanine Nucleotide Exchange Factor; rap GTP-Binding Proteins; Ras protein; ras Proteins - genetics; ras Proteins - physiology; Signal Transduction
• ISSN: 0950-9232; 1476-5594
• DOI: 10.1038/sj.onc.1201473

Ral GDP dissociation stimulator (RalGDS), a putative effector protein of Ras, stimulated the GDP/GTP exchange reaction of the post-tanslationally lipid-modified but not the unmodified form of Ral in response to epidermal growth factor in COS cells. The RalGDS action on Ral was enhanced by an active form of Ras but not a Ras mutant which was not post-translationally modified in the cells. The RalGDS activity was inhibited by acidic membrane phospholipids such as phosphatidylinositol and phosphatidylserine but not by phosphatidylcholine or phosphatidylethanolamine in vitro. The post-translationally modified form but not unmodified form of Ras, Ral, and Rap were incorporated in liposomes consisting of these phospholipids. When Ral was incorporated alone in the liposomes, RalGDS did not stimulate the dissociation of GDP from Ral. When Ral was incorporated with the GTP-bound form of Ras in the liposomes, RalGDS stimulated the dissociation of GDP from Ral, while the GDP-bound form of Ras did not affect the RalGDS action. The Ras-dependent Ral activation through RalGDS required the Ras-binding domain of RalGDS. Rap, which shared the same effector loop as Ras, also stimulated the dissociation of GDP from Ral through RalGDS in the liposomes, although Rap did not enhance the RalGDS action in COS cells. Taken together with our previous observations that Ras recruits RalGDS to the membrane, these results indicate that the post-translational modifications of Ras and Ral are important for Ras-dependent Ral activation through RalGDS and that colocalization of Ras and Ral on the membrane is necessary for Ral activation in intact cells.