Abstract
Several deletion mutants of Raf-1 were expressed with v- ras p21 or 14-3-3 protein in COS-7 cells and Sf9 cells and the interaction of Raf-1 with ras p21 or with 14-3-3 protein in intact cells was examined. Raf(1–135) (residues 1–135) and Raf(1–322) interacted with v- ras p21, but other deletion mutants such as Raf(136–322) or Raf(321–648) did not. Raf(1–322) interacted with 14-3-3 protein much more efficiently than Raf(321–648) did. While Raf(1–135) did not interact with 14-3-3 protein, Raf(136–322) did. These results clearly indicate that Raf-1 simultaneously interacts with both ras p21 and 14-3-3 protein through the distinct binding domains in intact cells.
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