Differential effects of Ras signaling through NFκB on skeletal myogenesis

Oncogenic Ras (H-Ras G12V) inhibits skeletal myogenesis through multiple signaling pathways. Previously, we demonstrated that the major downstream effecters of Ras (i.e., MEK/MAPK, RalGDS and Rac/Rho) play a minor, if any, role in the differentiation-defective phenotype of Ras myoblasts, Recently, NF kappaB, another Ras signaling target, has been shown to inhibit myogenesis presumably by stimulating cyclin D1 accumulation and cell cycle progression. In this study, we address the involvement of NF kappaB activation in the Ras-induced inhibition of myogenesis, Using H-Ras G12V and three G12V effector-loop variants, we detect high levels of NF kappaB transcriptional activity in C3H10T1/2-MyoD cells treated with differentiation medium. Myogenesis is blocked by all Ras proteins tested, yet only in the case of H-Ras G12V are cyclin D1 levels increased and cell cycle progression maintained, Expression of I kappaB alpha SR, an inhibitor of NF kappaB, does not reverse the differentiation-defective phenotype of Ras expressing cultures, but does induce differentiation in cultures treated with tumor necrosis factor (TNF alpha) or in cultures expressing the RelA/p65 subunit of NF kappaB, These data confirm that NF kappaB is a target of Ras and suggest that the cellular actions of NF kappaB require additional signals that are discriminated by the Ras effector-loop variants. Results with I kappaB alpha SR convincingly demonstrate that H-Ras G12V does not rely on NF kappaB activity to block myogenesis, an observation that continues to implicate another unidentified signaling pathway(s) in the inhibition of skeletal myogenesis by Ras.