Osmium tetroxide as a probe of RNA structure.
Osmium tetroxide as a probe of RNA structure.
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四氧化锇作为 RNA 结构的探针。
DOI：
10.1261/rna.057539.116
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发表时间：
2017-04
期刊：
影响因子：
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通讯作者：
Murchie AI
中科院分区：
文献类型：
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作者：
Zhang J;Li D;Zhang J;Chen D;Murchie AI
结构化RNA在细胞功能中具有核心作用。结构化RNA采用固定结构或经历动态构象变化的能力有助于它们在基因表达调控中的不同作用。尽管已经开发了许多生物物理和生物化学工具来研究结构化RNA，但仍然需要开发用于研究RNA结构的新方法，特别是允许在接近其天然细胞条件的溶液中研究RNA结构的方法。在这里，我们使用四氧化锇（OsO4）作为RNA结构的化学探针。在该方法中，我们使用基于荧光的测序技术来检测OsO4修饰的RNA。我们的特点是OsO4修饰的RNA的要求，通过调查三个已知的结构RNA：M盒，甘氨酸核糖开关RNA，和tRNAasp。我们的研究结果表明，OsO4主要修改RNA的尿嘧啶的构象暴露在表面上的RNA。我们还表明，OsO4的反应性在灵活的位置在RNA中的变化与配体驱动的RNA结构的构象变化。四氧化锇修饰的RNA将提供深入了解RNA的结构特征，是相关的其潜在的生物学功能。
Structured RNAs have a central role in cellular function. The capability of structured RNAs to adopt fixed architectural structures or undergo dynamic conformational changes contributes to their diverse role in the regulation of gene expression. Although numerous biophysical and biochemical tools have been developed to study structured RNAs, there is a continuing need for the development of new methods for the investigation of RNA structures, especially methods that allow RNA structure to be studied in solution close to its native cellular conditions. Here we use osmium tetroxide (OsO4) as a chemical probe of RNA structure. In this method, we have used fluorescence-based sequencing technologies to detect OsO4 modified RNA. We characterized the requirements for OsO4 modification of RNA by investigating three known structured RNAs: the M-box, glycine riboswitch RNAs, and tRNAasp. Our results show that OsO4 predominantly modifies RNA at uracils that are conformationally exposed on the surface of the RNA. We also show that changes in OsO4 reactivity at flexible positions in the RNA correlate with ligand-driven conformational changes in the RNA structure. Osmium tetroxide modification of RNA will provide insights into the structural features of RNAs that are relevant to their underlying biological functions.