The core enhancer is essential for proper timing of MyoD activation in limb buds and branchial arches

Abstract

Transgenic analyses have defined two transcriptional enhancers that regulate MyoD expression in mammals, the core enhancer and distal regulatory region; these enhancers exhibit complementary activities and together are sufficient to recapitulate MyoD expression in developing and mature skeletal muscle. The core enhancer is activated in presumptive muscle cells and determined myoblasts, suggesting an important role in initiating MyoD expression. Here, targeted mutagenesis in the mouse is used to identify necessary and redundant core enhancer functions. The core enhancer is essential for the timely initiation of MyoD expression in limb buds and branchial arches, as enhancer deletion delayed MyoD activation by 1 to 2 days in these muscle lineages. Functionally, this delay in MyoD transcription delayed the onset of muscle differentiation, as assayed by expression of the gene encoding for the early differentiation marker, Myogenin. In addition to these lineage-specific defects, a generalized, modest reduction in MyoD expression was observed in all muscle lineages and at all embryonic stages examined. Interestingly, however, a specific defect was not observed in the nascent myocytes at the medial and lateral aspects of the myotome, suggesting the existence of at least one other enhancer with this specificity. The core enhancer was also dispensable for Myf-5- and Pax-3-dependent regulation of MyoD transcription. These data demonstrate a differential requirement for core enhancer activity in muscle lineages derived from migratory precursors and suggest redundancy in cis regulatory mechanisms controlling myotomal MyoD expression.