Protein kinase C delta activates the MEK-ERK pathway in a manner independent of Ras and dependent on Raf

Although the involvement of protein kinase C (PKC) in the activation of the mitogen-activated protein (MAP) kinase pathway has been implicated through experiments using 12-O-tetradecanoylphorbol-13-acetate (TPA), there has been no direct demonstration that PKC activates the MAP kinase pathway, A Raf-dependent intact cell assay system for monitoring the activation of MAPK/ERK kinase (MEK) and extracellular signal-related kinase (ERK) permitted us to evaluate the role of PRC isotypes in MAP kinase activation. Treatment of cells with TPA or epidermal growth factor resulted in the activation of MEK and ERK. The activation of the MAP kinase pathway triggered by epidermal growth factor was completely inhibited by dominant-negative Ras (RasN17), whereas the activation triggered by TPA was not, consistent with previous observations. The introduction of an activated point mutant of PKC delta, but not PKC alpha or PKC epsilon, resulted in the activation of the MAP kinase pathway. The activation of MEK and ERK by an activated form of PKC delta requires the presence of c-Raf and is independent of RasN17. These results demonstrate that activation of PKC delta is sufficient for the activation of MER and ERK and that the pathway operates in a manner dependent on c-Raf and independent of Ras.