[No authors listed]
Calpain 3 (Capn3), a skeletal muscle-specific member of the calpain family, executes some non-proteolytic functions besides its role as a Ca2+-regulated proteolytic enzyme. Previously, we found that changes in Capn3 expression were linearly correlated with the degree of muscular atrophy following reversible sciatic nerve injury and that knockdown of Capn3 gene expression promoted myoblast differentiation. While the regulation of capn3 gene expression is interesting, transcriptional regulation of Capn3 is still unclear. In the present study, we provided experimental data showing that the myogenic enhancer factor 2A (MEF2A) regulated Capn3 gene expression. Firstly, the luciferase reporter assay and EMSA were performed and showed that ectopic expression of the Mef2a gene could bind to the predicted site of the Capn3 promoter region. Furthermore, in the L6 myoblast differentiation model in vitro, Capn3 gene expression was shown to be positively associated with the level of Mef2a by qRT-PCR, western-blotting, and immunocytochemistry. The Capn3 protein level decreased as MEF2A decreased when induced by Mef2a siRNA transfection in L6 myoblasts. Finally, the results of ChIP indicated that MEF2A occupied the promoter region of the Capn3 gene in rat denervated gastrocnemius muscle tissue. Based on these results, we proposed that MEF2A is a transcriptional regulator for Capn3 gene expression.
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