RKIP regulates MAP kinase signaling in cells with defective B-Raf activity

MAP kinase (MAPK) signaling results from activation of Raf kinases in response to external or internal stimuli. Here, we demonstrate that Raf kinase inhibitory protein (RKIP) regulates the activation of MAPK when B-Raf signaling is defective. We used multiple models including mouse embryonic fibroblasts (MEFs) and primary keratinocytes from RKIP- or Raf-deficient mice as well as allografts in mice to investigate the mechanism. Loss of B-Raf protein or activity significantly reduces MAPK activation in these cells. We show that RKIP depletion can rescue the compromised ERK activation and promote proliferation, and this rescue occurs through a Raf-1 dependent mechanism. These results provide formal evidence that RKIP is a bona fide regulator of Raf-1. We propose a new model in which RKIP plays a key role in regulating the ability of cells to signal through Raf-1 to ERK in B-Raf compromised cells.

► Generated RKIP-/- and RKIP-/-B-Raf-/- mice to measure effect of RKIP on MAPK.
► B-Raf but not RKIP deficiency or inactivation compromises ERK signaling.
► RKIP loss rescues ERK activity in B-Raf-deficient MEFs and primary keratinocytes.
► RKIP-dependent rescue of ERK in B-Raf deficient cells is Raf-1 dependent.
► RKIP loss rescues cell growth in B-Raf -/- MEFs in culture and mouse allografts.