Abstract

N ⁶-methyladenosine (m⁶A) plays an essential role in regulating gene expression. However, the effect of m⁶A on skeletal myoblast differentiation and the underlying mechanisms are still unclear. Here, we ascertained mRNA m⁶A methylation exhibited declined changes during bovine skeletal myoblast differentiation, and both MEF2C mRNA expression and m⁶A levels were significantly increased during myoblast differentiation. We found that MEF2C with mutated m⁶A sites significantly inhibited myoblast differentiation compared with wild-type MEF2C. METTL3 promoted MEF2C protein expression through posttranscriptional modification in an m⁶A-YTHDF1-dependent manner. Moreover, MEF2C promoted the expression of METTL3 by binding to its promoter. These results revealed that there is a positive feedback loop between these molecules in myoblast differentiation. Our study provided new insights into skeletal muscle differentiation and fusion, which may provide an RNA methylation-based approach for molecular genetics and breeding in livestock as well as for the treatment of muscle-related diseases.