cAMP-dependent protein kinase represses myogenic differentiation and the activity of the muscle-specific helix-loop-helix transcription factors Myf-5 and MyoD.

Myf-5 and MyoD are members of a family of muscle-specific basic helix-loop-helix (bHLH) proteins that are fundamental for myogenic cell differentiation and transcriptional activation of muscle-specific genes. Here we report that elevated levels of the intracellular signaling molecule cAMP and overexpression of cAMP-dependent protein kinase (PKA) inhibit myogenic differentiation. PKA represses the transcriptional activation of muscle-specific genes by the myogenic regulators Myf-5 and MyoD. The repression is directed at the basic HLH domain and is mediated through the E-box DNA consensus motif to which these proteins bind. However, phosphorylation of Myf-5 and MyoD by PKA in vitro does not affect their ability to bind to DNA. PKA specifically inhibits the activity of myogenic bHLH proteins, but not of other HLH proteins, such as the ubiquitously expressed E2A gene products E12 and E47 (E2-5). Our results demonstrate that PKA mediates the cAMP-induced inhibition of muscle cell differentiation by repressing the activity of Myf-5 and MyoD. The inhibition by PKA occurs post-translationally and presumably affects the transactivation process at a step following DNA-binding. The regulation of Myf-5 and MyoD function by a cAMP-dependent pathway may partly explain how external signals generated by serum and certain peptide growth factors can be transduced to the nucleus and inhibit dominant-acting factors that are responsible for myoblast differentiation.