m6 A Methylases Regulate Myoblast Proliferation, Apoptosis and Differentiation

Abstract

N6-methyladenosine (m6 A) plays an important role in regulating gene expression. Previous studies found that m6 A methylation affects skeletal muscle development. However, the effect of m6 A methylases on bovine skeletal myogenesis is still unclear. Here, we found that the expression of m6 A demethylases (FTO and ALKBH5) was significantly higher in the longissimus dorsi muscle of adult cattle than in newborn cattle. In contrast, the expression of m6 A methyltransferases (METTL3, METTL14 and WTAP) was reduced. The mRNA expression of all five genes was found to be increased during the myogenesis of myoblasts in vitro. Knockdown of FTO or METTL3 promoted myoblast proliferation, inhibited myoblast apoptosis and suppressed myogenic differentiation, whereas ALKBH5 knockdown had the opposite effect. METTL14 knockdown enhanced myoblast proliferation and impaired myogenic differentiation. WTAP knockdown attenuated proliferation and contributed to apoptosis but did not affect differentiation. Furthermore, the functional domains of these five m6 A methylases are conserved across species. Our results suggest that m6 A methylases are involved in regulating skeletal muscle development and that there may be a complex network of m6 A methylation regulating skeletal myogenesis.