Identification of residues in the cysteine-rich domain of Raf-1 that control Ras binding and Raf-1 activity.

摘要: We have identified mutations in Raf-1 that increase binding to Ras. The were making use of three mutant forms Ras reduced (Winkler, D. G., Johnson, J. C., Cooper, A., and Vojtek, A. B. (1997) Biol. Chem. 272, 24402–24409). One mutation Raf-1, N64L, suppresses the R41Q but not other mutants, suggesting this structurally complements mutation. Missense substitutions residues 143 144 cysteine-rich domain isolated multiple times. These R143Q, R143W, K144E, general suppressors different mutants had increased interaction with non-mutant Each was slightly activated relative wild-type a transformation assay. In addition, two R143W active when tested for induction germinal vesicle breakdown Xenopus oocytes. Interestingly, all ability N-terminal regulatory region inhibit oocyte induced by C-terminal catalytic Raf-1. propose direct or indirect between N- regions is K144E mutations, thereby increasing access Ras-binding activity.