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首页> 外文期刊>Biochimica et Biophysica Acta. Gene Regulatory Mechanisms >Hydroxymethylation of DNA influences nucleosomal conformation and stability in vitro
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Hydroxymethylation of DNA influences nucleosomal conformation and stability in vitro

机译：DNA的羟甲基化影响体外的核小体构象和稳定性

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Background: Hydroxymethylation of DNA at the C5 position of cytosine (5hmC) is recognized as an important epigenetic mark. The molecular role of 5hmC in gene regulation, however, is not well understood. Methods: We studied the effects of 5-hydroxymethylation (5hmC) on nucleosome properties in vitro using a combination of biochemical and fluorescence assays. Competitive reconstitution was used to evaluate the effect of 5hmC on nucleosome formation. The effects of 5hmC on nucleosome compactness and stability were characterized using FRET assays. These findings have also been compared with another important epigenetic mark, the cytosine methylation (5mC) of DNA. Results: We observed that hydroxymethylation increases the binding affinity of DNA for the histone octamer. The formed nucleosome exhibits slightly different conformations based on the sequence and epigenetic context of DNA. DNA hydroxymethylation decreases the stability of formed nucleosomes in salt-induced dissociation processes. Conclusion: DNA containing 5hmC is more likely to be incorporated into nucleosomes. Once formed, the 5hmC nucleosomes might be in an open and transcriptionally active state due to the weakened interaction of hydroxymethylated DNA with the H2A-H2B dimers. General significance: Our results reveal the effect of 5hmC on regulating nucleosome compactness and stability in vitro.
机译：背景：胞嘧啶（5hmC）C5位置的DNA羟甲基化被认为是重要的表观遗传标记。然而，人们对5hmC在基因调控中的分子作用还知之甚少。方法：我们结合生化和荧光分析研究了5-羟甲基化（5hmC）对体外核小体性质的影响。竞争性重建用于评估5hmC对核小体形成的影响。使用FRET分析表征了5hmC对核小体紧密性和稳定性的影响。这些发现也已与另一个重要的表观遗传标记DNA的胞嘧啶甲基化（5mC）相比较。结果：我们观察到羟甲基化增加了DNA对组蛋白八聚体的结合亲和力。基于DNA的序列和表观遗传背景，形成的核小体展示出稍微不同的构象。 DNA羟甲基化会降低盐诱导的解离过程中形成的核小体的稳定性。结论：含有5hmC的DNA更有可能掺入核小体中。 5hmC核小体一旦形成，由于羟甲基化DNA与H2A-H2B二聚体的相互作用减弱，可能处于开放和转录活性状态。一般意义：我们的结果揭示了5hmC对体外调节核小体紧密性和稳定性的影响。

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