Myocyte Enhancer Factor 2C and Myogenin Up-regulate Each Other's Expression and Induce the Development of Skeletal Muscle in P19 Cells

Two families of transcription factors, myogenic regulatory factors (MRFs) and myocyte enhancer factor 2 (MEF2), function synergistically to regulate myogenesis. In addition to activating structural muscle-specific genes, MRFs and MEF2 activate each other's expression. The MRF, myogenin, can activate MEF2 DNA binding activity when transfected into fibroblasts and, in turn, the myogenin promoter contains essential MEF2 DNA binding elements. To determine which MEF2 is involved in this regulation, P19 cells stably expressing MyoD and myogenin were compared for their ability to activate the expression of MEF2 family members. There was very little cross-activation of MyoD expression by myogenin and vice versa. Myogenin expression, and not MyoD, was found to up-regulate MEF2C expression. MEF2A, -B, and -D expression levels were not up-regulated by overexpression of either MyoD or myogenin. To examine whether MEF2C can differentially regulate MyoD or myogenin expression, P19 cell lines overexpressing MEF2C were analyzed. MEF2C induced myogenesis in P19 cells and up-regulated the expression of myogenin with 25-fold greater efficiency than that of MyoD. Therefore, myogenin and MEF2C participate in a regulatory loop in differentiating stem cells. This positive regulation does not extend to MyoD or the other MEF2 family members. Consequently, MEF2C appears to play a specific role in early events of myogenesis.