Structural analysis of RNA in living cells by in vivo synchrotron X-ray footprinting.
Structural analysis of RNA in living cells by in vivo synchrotron X-ray footprinting.
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DOI：
10.1016/s0076-6879(09)68012-5
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发表时间：
2009
影响因子：
--
通讯作者：
Woodson SA
中科院分区：
文献类型：
--
作者：
Adilakshmi T;Soper SF;Woodson SA
化学足迹法被广泛用于探测核酸及其复合物的溶液结构。在许多可用的修饰试剂中，羟基自由基在其提供关于核酸骨架的溶剂可及性的核苷酸水平信息的能力方面是例外的。直到最近，羟基自由基足迹一直局限于体外实验。我们描述了使用同步加速器X射线辐射在细胞内产生羟基自由基的RNA-蛋白质复合物在体内的有效足迹。该技术给出的结果与体外足迹实验一致，差异反映了体内RNA的明显结构变化。
Chemical footprinting methods are widely used to probe the solution structures of nucleic acids and their complexes. Among the many available modifying reagents, hydroxyl radical is exceptional in its ability to provide nucleotide-level information on the solvent accessibility of the nucleic acid backbone. Until recently, hydroxyl radical footprinting has been limited to in vitro experiments. We describe the use of synchrotron X-radiation to generate hydroxyl radicals within cells for effective footprinting of RNA-protein complexes in vivo. This technique gives results that are consistent with in vitro footprinting experiments, with differences reflecting apparent structural changes to the RNA in vivo.