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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Conformational flexibility in the chromatin remodeler RSC observed by electron microscopy and the orthogonal tilt reconstruction method
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Conformational flexibility in the chromatin remodeler RSC observed by electron microscopy and the orthogonal tilt reconstruction method

机译：用电子显微镜和正交倾斜重建方法观察染色质重塑剂RSC中的构象柔性

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摘要

Chromatin remodeling complexes (remodelers) are large, multi-subunit macromolecular assemblies that use ATP hydrolysis to alter the structure and positioning of nucleosomes. The mechanisms proposed for remodeler action on nucleosomes are diverse, and require structural evaluation and insights. Previous reconstructions of remodelers using electron microscopy revealed interesting features, but also significant discrepancies, prompting new approaches. Here, we use the orthogonal tilt reconstruction method, which is well suited for heterogeneous samples, to provide a reconstruction of the yeast RSC (remodel the structure of chroma-tin) complex. Two interesting features are revealed: first, we observe a deep central cavity within RSC, displaying a remarkable surface complementarity for the nucleosome. Second, we are able to visualize two distinct RSC conformers, revealing a major con-formational change in a large protein "arm," which may shift to further envelop the nucleosome. We present a model of the RSC-nucleosome complex that rationalizes the single molecule results obtained by using optical tweezers and also discuss the mechanistic implications of our structures.
机译：染色质重塑复合物（重塑剂）是大型的多亚基大分子装配体，可使用ATP水解来改变核小体的结构和位置。拟议的重塑对核小体作用的机制多种多样，需要结构评估和见解。以前使用电子显微镜对重构者进行的重建显示出有趣的功能，但也存在重大差异，从而催生了新的方法。在这里，我们使用非常适合异质样品的正交倾斜重建方法，来提供酵母RSC（重塑色氨酸的结构）复合物的重建。揭示了两个有趣的特征：首先，我们观察到RSC内的深中心腔，显示出核小体的显着表面互补性。其次，我们能够可视化两个不同的RSC构象异构体，揭示了大型蛋白质“臂”中的主要构象变化，该变化可能会进一步包裹核小体。我们提出了一个RSC-核小体复合物模型，该模型合理化了使用光镊获得的单分子结果，并讨论了我们结构的机械含义。

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