Differential regulation of rasGAP and neurofibromatosis gene product activities

Abstract

THE ras-encoded p21^ras proteins bind GTP very tightly, but catalyse hydrolysis to GDP very slowly¹. In humans, two genes encode proteins that stimulate this GTPase activity (GAP, or GTPase-activating proteins)², one of relative molecular mass 120,000, referred to as p120-GAP, and another NF1-GAP, which is encoded by the neurofibromatosis type-1 gene^3–5. Both GAPs are widely expressed in mammalian tissues^6,7. Here we show that although they will both bind oncogenic mutants of p21^ras, neither will stimulate their GTPase activity. NF1-GAP binds to the p21^ras proteins up to 300 times more efficiently than p120-GAP. The two GAPs are inhibited to different extents by certain lipids: micromolar concentrations of arachidonate, phosphatidate and phosphatidyIinositol-4,5-bisphosphate affect only NF1-GAP. This inhibition does not compete with p21^ras, and lipid-inactivated NF1-G AP can still bind p21^ras. We used the detergent dodecyl maltoside, which inhibits only NF1-GAP, to distinguish between the two activities in cell extracts and found both types present together in several mammalian cell lines. In contrast, GAP activity in extracts of Xenopus oocytes was not affected by dodecyl maltoside. By these criteria, the mammalian cells contain both GAP activities and the oocytes have only p120-like GAP activity. These results indicate that more than one GAP regulates p21^ras in the same cell.