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Published on: April 29, 2011
Genome-wide mapping of the binding sites of myocyte enhancer factor 2A in chicken primary myoblasts
Xinglong Wang1, Jiannan Zhang1, Jiancheng Su1
1Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, PR China.
Abstract:
Myocyte enhancer factor 2A (MEF2A) is a transcription factor that plays a critical role in cell proliferation, differentiation and apoptosis. In contrast to the wide characterization of its regulation mechanism in mammalian skeletal muscle, its role in chickens is limited. Especially, its wide target genes remain to be identified. Therefore, we utilized Cleavage Under Targets and Tagmentation (CUT&Tag) technology to reveal the genome-wide binding profile of MEF2A in chicken primary myoblasts thus gaining insights into its potential role in muscle development. Our results revealed that MEF2A binding sites were primarily distributed in intergenic and intronic regions. Within the promoter region, although only 8.87% of MEF2A binding sites were found, these binding sites were concentrated around the transcription start site (TSS). Following peak annotation, a total of 1903 genes were identified as potential targets of MEF2A. Gene Ontology (GO) enrichment analysis further revealed that MEF2A target genes may be involved in the regulation of embryonic development in multiple organ systems, including muscle development, gland development, and visual system development. Moreover, a comparison of the MEF2A target genes identified in chicken primary myoblasts with those in mouse C2C12 cells revealed 388 target genes are conserved across species, 1515 target genes are chicken specific. Among these conserved genes, ankyrin repeat and SOCS box containing 5 (ASB5), transmembrane protein 182 (TMEM182), myomesin 2 (MYOM2), leucyl and cystinyl aminopeptidase (LNPEP), actinin alpha 2 (ACTN2), sorbin and SH3 domain containing 1 (SORBS1), ankyrin 3 (ANK3), sarcoglycan delta (SGCD), and ORAI calcium release-activated calcium modulator 1 (ORAI1) exhibited consistent expression patterns with MEF2A during embryonic muscle development. Finally, TMEM182, as an important negative regulator of muscle development, has been validated to be regulated by MEF2A by dual-luciferase and quantitative real-time PCR (qPCR) assays. In summary, our study for the first time provides a wide landscape of MEF2A target genes in chicken primary myoblasts, which supports the active role of MEF2A in chicken muscle development.
Insights
Myocyte enhancer factor 2A (MEF2A) is crucial for chicken muscle development. This study mapped MEF2A
Area of Science:
- Molecular Biology
- Developmental Biology
- Genomics
Background:
- Myocyte enhancer factor 2A (MEF2A) is a key transcription factor regulating muscle development in mammals.
- Its specific functions and target genes in avian species, particularly chickens, remain largely uncharacterized.
- Understanding MEF2A's role is essential for advancing knowledge of avian muscle development.
Purpose of the Study:
- To elucidate the genome-wide binding profile of MEF2A in chicken primary myoblasts.
- To identify novel MEF2A target genes and understand their function in chicken muscle development.
- To compare MEF2A targets between chicken and mouse models to identify conserved and species-specific regulatory networks.
Main Methods:
- Cleavage Under Targets and Tagmentation (CUT&Tag) technology was employed to map MEF2A binding sites genome-wide.
- Bioinformatic analyses, including peak annotation and Gene Ontology (GO) enrichment, were performed on identified binding sites.
- Comparative genomics and molecular assays (dual-luciferase, qPCR) were used to validate target genes and their regulatory relationships.
Main Results:
- MEF2A binding predominantly occurred in intergenic and intronic regions, with a concentration near transcription start sites in promoters.
- A total of 1903 potential MEF2A target genes were identified, implicated in embryonic development across multiple organ systems.
- 388 MEF2A target genes were conserved between chicken and mouse, while 1515 were chicken-specific, with several key muscle-related genes showing conserved expression patterns.
Conclusions:
- This study provides the first comprehensive map of MEF2A target genes in chicken primary myoblasts.
- MEF2A plays a significant role in chicken muscle development, regulating a broad spectrum of genes involved in embryonic development.
- The findings offer valuable insights into the conserved and species-specific mechanisms of MEF2A-mediated gene regulation in vertebrates.
