Cryo-EM in the study of challenging systems: the human transcription pre-initiation complex.

Cryo-EM in the study of challenging systems: the human transcription pre-initiation complex.
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DOI:
10.1016/j.sbi.2016.09.009
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发表时间:
2016-10
影响因子:
6.8
通讯作者:
He, Yuan
He, Yuan
中科院分区:
生物学2区
文献类型:
--
作者:
Nogales, Eva;Louder, Robert K.;He, Yuan

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单粒子低温电子显微镜(cryo-EM)是一种无需结晶即可对大分子进行结构表征的技术。对于理想地适合于冷冻EM研究的某些类型的样品,已经可以按照相对标准的程序达到高分辨率结构。其他生物系统仍然具有高度挑战性,即使是冷冻EM。挑战可能涉及样品的稀缺性,复合物的稳定性差,以及最常见的生物分子的固有灵活性。在这些具有挑战性的样品中,有大的真核转录复合物,它们具有所有这些缺点。在这里,我们报告我们最近如何试图克服这些挑战,以提高我们的结构理解的人类转录前起始复合物组装和转录起始过程。平行的努力也进行了芽殖酵母转录起始复合物,允许比较,建立两个系统之间的整体保护和特定的结构差异。
Single particle cryo-Electron Microscopy (cryo-EM) is a technique that allows the structural characterization of macromolecules without the need for crystallization. For certain type of samples that are ideally suited for cryo-EM studies it has been possible to reach high-resolution structures following relatively standard procedures. Other biological systems remain highly challenging, even for cryo-EM. Challenges may involve the scarcity of the sample, poor stability of the complexes, and most often, the intrinsic flexibility of biological molecules. Among these challenging samples are large eukaryotic transcription complexes, which suffer from all such shortcomings. Here we report how we have recently tried to overcome those challenges in order to improve our structural understanding of the human transcription pre-initiation complex assembly and the transcription initiation process. Parallel efforts have also been carried out for budding yeast transcription initiation complexes, allowing comparisons that establish both the overall conservation and the specific structural differences between the two systems.
电子计数和束诱导运动校正可实现近原子分辨率的单粒子冷冻电镜。
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