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首页> 外文期刊>Nucleic acids research >Phase-plate cryo-EM structure of the Widom 601 CENP-A nucleosome core particle reveals differential flexibility of the DNA ends
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Phase-plate cryo-EM structure of the Widom 601 CENP-A nucleosome core particle reveals differential flexibility of the DNA ends

机译：WIDOM 601 CENP-核心核心粒子的相板冷冻 - EM结构揭示了DNA末端的差异柔韧性

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The histone H3 variant CENP-A marks centromeres epigenetically and is essential for mitotic fidelity. Previous crystallographic studies of the CENP-A nucleosome core particle (NCP) reconstituted with a human α-satellite DNA derivative revealed both DNA ends to be highly flexible, a feature important for CENP-A mitotic functions. However, recent cryo-EM studies of CENP-A NCP complexes comprising primarily Widom 601 DNA reported well-ordered DNA ends. Here, we report the cryo-EM structure of the CENP-A 601 NCP determined by Volta phase-plate imaging. The data reveal that one (‘left’) 601 DNA end is well ordered whereas the other (‘right’) end is flexible and partly detached from the histone core, suggesting sequence-dependent dynamics of the DNA termini. Indeed, a molecular dynamics simulation of the CENP-A 601 NCP confirmed the distinct dynamics of the two DNA extremities. Reprocessing the image data using two-fold symmetry yielded a cryo-EM map in which both DNA ends appeared well ordered, indicating that such an artefact may inadvertently arise if NCP asymmetry is lost during image processing. These findings enhance our understanding of the dynamic features that discriminate CENP-A from H3 nucleosomes by revealing that DNA end flexibility can be fine-tuned in a sequence-dependent manner.
机译：组蛋白H3变体CENP-A标记焦粒表征，对有丝分裂保真来说是必不可少的。以人α-卫星DNA衍生物重构的CENP-A核心颗粒（NCP）的先前结晶研究显示，两种DNA末端都是高度柔韧的，该特征对于CENP-A有丝分子功能很重要。然而，最近CENP-A NCP复合物的Cryo-EM研究，主要包括Widom 601 DNA报告了有序的DNA末端。这里，我们报告了由Volta相板成像确定的CENP-A 601 NCP的Cryo-EM结构。数据显示，一个（'左'）601 DNA端是良好的订购，而另一个（'右'）末端是柔性的，并且部分地从组蛋白核心脱离，表明DNA末端的序列依赖性动态。实际上，CENP-A 601 NCP的分子动力学模拟证实了两个DNA四肢的不同动态。使用双倍对称进行重新处理图像数据产生了一种低温 - EM图，其中两个DNA结束出现良好的顺序，表明如果在图像处理期间丢失NCP不对称，则可能无意中出现这种人工制品。这些发现通过揭示通过序列依赖性方式可以微调DNA最终柔性来增强鉴别H3核体的动态特征的理解。

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