Structural analysis and cellular visualization of APP RNA G-quadruplex
Structural analysis and cellular visualization of APP RNA G-quadruplex
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APP RNA G-四联体的结构分析和细胞可视化
DOI：
10.1039/c9sc02768h
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发表时间：
2019-12-28
期刊：
影响因子：
8.4
通讯作者：
Kwok, Chun Kit
中科院分区：
文献类型：
--
作者：
Lyu, Kaixin;Chen, Shuo-Bin;Kwok, Chun Kit
RNA G-四链体（RG 4）是一种新兴的结构基序，在化学和生物学中具有关键重要性;然而，目前RG 4的结构信息有限，其在细胞中的折叠状态和功能仍然难以捉摸。在这里，我们开发并采用多学科的方法来表征体外和细胞中感兴趣的单个rG 4的结构，形成和功能。我们将这种策略应用于淀粉样前体蛋白（APP）转录中的生物学重要的RG 4，并揭示APP RG 4的独特结构特征。值得注意的是，我们可视化的APP RG 4在细胞中使用APP特异性G-四链体触发的荧光杂交（GTFH）探针的形成，并报告APP RG 4在翻译中的调节作用依赖于RG 4的热稳定性，提供证据的存在和稳定的RG 4结构在基因调控中的意义。
RNA G-quadruplexes (rG4s) are emerging structural motifs that are of pivotal importance in chemistry and biology; however, the current structural information of rG4s is limited, with their folding status and functions in cells remaining elusive. Here, we develop and employ a multi-disciplinary approach to characterize the structure, formation and function of an individual rG4 of interest in vitro and in cells. We apply this strategy to a biologically important rG4 in amyloid precursor protein (APP) transcript and reveal distinct structural features of APP rG4. Notably, we visualize the formation of APP rG4 in cells using an APP-specific G-quadruplex-triggered fluorogenic hybridization (GTFH) probe and report that the regulatory role of APP rG4 in translation is dependent on rG4 thermostability, providing evidence to the existence and significance of the stable rG4 structure in gene regulation.