Sequential association of myogenic regulatory factors and E proteins at muscle-specific genes.
Sequential association of myogenic regulatory factors and E proteins at muscle-specific genes.
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DOI：
10.1186/2044-5040-1-14
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发表时间：
2011-04-04
期刊：
影响因子：
4.9
通讯作者：
Davie JK
中科院分区：
文献类型：
--
作者：
Londhe P;Davie JK
骨骼肌中的基因表达由一个碱性螺旋-环-螺旋转录因子家族控制，该家族被称为成肌调节因子（MRF）。MRF与E蛋白一起调节肌发生过程中的基因表达。然而，MRF激活基因表达的确切机制尚不清楚。在这项工作中，我们试图确定MRF和E蛋白在整个分化过程中对肌肉特异性基因的结合谱。我们在C2 C12分化的时间过程中对肌细胞生成素、MyoD、Myf 5和E蛋白进行了染色质免疫沉淀（ChIP）测定，得到了几个令人惊讶的发现。募集模式对每个测试的启动子是特异性的。E蛋白的募集通常与MRF的到达一致，但结合谱与MRF结合谱并不完全重叠。我们发现E12/E47在增殖过程中与某些启动子结合，但每个测试的基因在分化过程中优先与HEB结合。我们还表明，MyoD，肌细胞生成素和Myf 5在肌肉分化过程中对这些启动子中的每一个都有瞬时作用。我们还发现，RNA聚合酶II占用与这些启动子的转录谱相关。ChIP测序分析证实MyoD、肌细胞生成素和Myf 5共占据启动子。我们的数据揭示了MyoD、肌细胞生成素、Myf 5和HEB在肌肉特异性启动子上的顺序关联。这些数据表明，每个MRF，包括Myf 5，都有助于这里分析的每个基因的基因表达。观察到的动态结合曲线表明，MRF和E蛋白被独立地募集到启动子。
Gene expression in skeletal muscle is controlled by a family of basic helix-loop-helix transcription factors known as the myogenic regulatory factors (MRFs). The MRFs work in conjunction with E proteins to regulate gene expression during myogenesis. However, the precise mechanism by which the MRFs activate gene expression is unclear. In this work, we sought to define the binding profiles of MRFs and E proteins on muscle-specific genes throughout a time course of differentiation. We performed chromatin immunoprecipitation (ChIP) assays for myogenin, MyoD, Myf5 and E proteins over a time course of C2C12 differentiation, resulting in several surprising findings. The pattern of recruitment is specific to each promoter tested. The recruitment of E proteins often coincides with the arrival of the MRFs, but the binding profile does not entirely overlap with the MRF binding profiles. We found that E12/E47 is bound to certain promoters during proliferation, but every gene tested is preferentially bound by HEB during differentiation. We also show that MyoD, myogenin and Myf5 have transient roles on each of these promoters during muscle differentiation. We also found that RNA polymerase II occupancy correlates with the transcription profile of these promoters. ChIP sequencing assays confirmed that MyoD, myogenin and Myf5 co-occupy promoters. Our data reveal the sequential association of MyoD, myogenin, Myf5 and HEB on muscle-specific promoters. These data suggest that each of the MRFs, including Myf5, contribute to gene expression at each of the geness analyzed here.. The dynamic binding profiles observed suggest that MRFs and E proteins are recruited independently to promoters.
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