Nucleobases Undergo Dynamic Rearrangements during RNA Tertiary Folding.
Nucleobases Undergo Dynamic Rearrangements during RNA Tertiary Folding.
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DOI：
10.1016/j.jmb.2016.09.015
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发表时间：
2016-11-06
影响因子：
5.6
通讯作者：
Hall, Kathleen B.
中科院分区：
文献类型：
--
作者：
Welty, Robb;Hall, Kathleen B.
在共晶体中，23 S rRNA的GTbR中心（GAC）的三级结构非常紧凑。它通过长程氢键和核碱基堆叠以及在其3向连接内形成的三环来稳定。其从二级结构到三级结构的折叠途径以前尚未观察到，但在平衡实验中显示需要Mg 2+离子。荧光核苷酸2-氨基嘌呤被取代在60个核苷酸的GAC内的选定位点。随着RNA折叠，在1 ms至100 s的时间过程中监测加入MgCl 2后的荧光强度变化。折叠途径在这里被揭示为通过几个中间体是分级的。对折叠过程中碱基的观察为研究折叠过程和途径提供了新的视角，揭示了折叠转变过程中碱基构象交换的动力学。
The tertiary structure of the GTPase Center (GAC) of 23S rRNA as seen in cocrystals is extremely compact. It is stabilized by long-range hydrogen bonds and nucleobase stacking, and a triloop that forms within its 3-way junction. Its folding pathway from secondary structure to tertiary structure has not been previously observed, but it was shown in equilibrium experiments to require Mg2+ ions. The fluorescent nucleotide 2-aminopurine was substituted at selected sites within the 60 nucleotide GAC. Fluorescence intensity changes upon addition of MgCl2 were monitored over a time-course from 1 ms to 100 s as the RNA folds. The folding pathway is revealed here to be hierarchical through several intermediates. Observation of the nucleobases during folding provides a new perspective on the process and the pathway, revealing the dynamics of nucleobase conformational exchange during the folding transitions.
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