Analysis of RNA structure using small-angle X-ray scattering
Analysis of RNA structure using small-angle X-ray scattering
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DOI：
10.1016/j.ymeth.2016.10.008
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发表时间：
2017-01-15
期刊：
影响因子：
4.8
通讯作者：
Musier-Forsyth, Karin
中科院分区：
文献类型：
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作者：
Cantara, William A.;Olson, Erik D.;Musier-Forsyth, Karin
氨酰-tRNA合成酶（阿尔斯）除了在将特定氨基酸正确连接到同源tRNA上的作用之外，还被发现具有许多替代功能，并且经常结合并作用于其他核酸。与tRNA明确的3D结构相反，aaRS识别的许多其他RNA的结构尚未得到解决。尽管在使用X射线晶体学（XRC），核磁共振（NMR）光谱和低温电子显微镜（cryoEM）的生物分子的结构表征的进步，解决RNA结构的重大挑战仍然存在。最近，小角X射线散射（SAXS）已越来越多地用于表征RNA和RNA-蛋白质复合物的三维结构。SAXS能够在生理条件下提供低分辨率的三级结构信息，并且与XRC，NMR和cryo-EM相比，样品制备和数据分析要求较低。在本文中，我们描述了RNA和RNA-蛋白质样品制备、SAXS数据收集、数据分析和结构模型构建过程中的最佳实践。(C)2016 Elsevier Inc. All rights reserved.
In addition to their role in correctly attaching specific amino acids to cognate tRNAs, aminoacyl-tRNA synthetases (aaRS) have been found to possess many alternative functions and often bind to and act on other nucleic acids. In contrast to the well-defined 3D structure of tRNA, the structures of many of the other RNAs recognized by aaRSs have not been solved. Despite advances in the use of X-ray crystallography (XRC), nuclear magnetic resonance (NMR) spectroscopy and cryo-electron microscopy (cryoEM) for structural characterization of biomolecules, significant challenges to solving RNA structures still exist. Recently, small-angle X-ray scattering (SAXS) has been increasingly employed to characterize the 3D structures of RNAs and RNA-protein complexes. SAXS is capable of providing low-resolution tertiary structure information under physiological conditions and with less intensive sample preparation and data analysis requirements than XRC, NMR and cryo-EM. In this article, we describe best practices involved in the process of RNA and RNA-protein sample preparation, SAXS data collection, data analysis, and structural model building. (C) 2016 Elsevier Inc. All rights reserved.