Genome-wide analysis of chromatin structure changes upon MyoD binding in proliferative myoblasts during the cell cycle
Genome-wide analysis of chromatin structure changes upon MyoD binding in proliferative myoblasts during the cell cycle
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细胞周期期间增殖性成肌细胞中 MyoD 结合后染色质结构变化的全基因组分析
DOI：
10.1093/jb/mvab001
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Ohkawa Yasuyuki
中科院分区：
文献类型：
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作者：
Wu Qianmei;Fujii Takeru;Harada Akihito;Tomimatsu Kosuke;Miyawaki-Kuwakado Atsuko;Fujita Masatoshi;Maehara Kazumitsu;Ohkawa Yasuyuki
MyoD是一种肌源性分化蛋白，已研究其在骨骼肌分化中的关键作用。表达MyoD的成肌细胞具有随着异位细胞增殖而分化的潜能。然而，鲜为人知的是，在增殖性成肌细胞中MyoD结合对染色质结构的影响。在这项研究中，我们通过染色质免疫沉淀测序评估了细胞周期中MyoD结合基因组区域周围的染色质结构。在增殖的小鼠C2 C12成肌细胞中，在细胞周期的三个阶段（G1，S，G2/M）期间进行组蛋白修饰的全基因组分析。我们发现，在细胞周期中，与MyoD未结合的基因组区域相比，MyoD结合的基因组区域具有较高水平的活性组蛋白修饰，如H3 K4 me 1/2/3和H3 K27 ac。我们还证明了在细胞周期期间维持升高的H3 K4 me 2/3修饰水平，而在后期阶段，H3 K27 ac和H3 K4 me 1修饰水平降低至与MyoD未结合的基因组区域相同的水平。免疫印迹分析显示，MyoD丰度在G1期高，然后在S和G2/M期下降。我们的研究结果表明，MyoD结合形成选择性的表观遗传记忆与H3 K4 me 2/3在细胞周期期间，除了通过活性染色质形成与转录耦合的肌原性基因诱导。
MyoD, a myogenic differentiation protein, has been studied for its critical role in skeletal muscle differentiation. MyoD-expressing myoblasts have a potency to be differentiated with proliferation of ectopic cells. However, little is known about the effect on chromatin structure of MyoD binding in proliferative myoblasts. In this study, we evaluated the chromatin structure around MyoD-bound genome regions during the cell cycle by chromatin immunoprecipitation sequencing. Genome-wide analysis of histone modifications was performed in proliferative mouse C2C12 myoblasts during three phases (G1, S, G2/M) of the cell cycle. We found that MyoD-bound genome regions had elevated levels of active histone modifications, such as H3K4me1/2/3 and H3K27ac, compared with MyoD-unbound genome regions during the cell cycle. We also demonstrated that the elevated H3K4me2/3 modification level was maintained during the cell cycle, whereas the H3K27ac and H3K4me1 modification levels decreased to the same level as MyoD-unbound genome regions during the later phases. Immunoblot analysis revealed that MyoD abundance was high in the G1 phase then decreased in the S and G2/M phases. Our results suggest that MyoD binding formed selective epigenetic memories with H3K4me2/3 during the cell cycle in addition to myogenic gene induction via active chromatin formation coupled with transcription.