Groove modification of siRNA duplexes to elucidate siRNA-protein interactions using 7-bromo-7-deazaadenosine and 3-bromo- 3-deazaadenosine as chemical probes
Groove modification of siRNA duplexes to elucidate siRNA-protein interactions using 7-bromo-7-deazaadenosine and 3-bromo- 3-deazaadenosine as chemical probes
复制标题
使用 7-溴-7-脱氮腺苷和 3-溴-3-脱氮腺苷作为化学探针对 siRNA 双链体进行凹槽修饰以阐明 siRNA-蛋白质相互作用
DOI：
10.1039/c6ob01866a
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Noriaki Minakawa
中科院分区：
文献类型：
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作者：
Noriko Saito-Tarashima;Hirotaka Kira;Tomoya Wada;Kazuya Miki;Shiho Ide;Naoshi Yamazaki;Akira Matsuda;Noriaki Minakawa
阐明RNA和蛋白质之间的动态相互作用对于理解由RNA调控的生物过程（例如RNA干扰（RNAi））是必不可少的。本研究以7-溴-7-脱氮腺苷（Br 7 C7 A）和3-溴-3-脱氮腺苷（Br 3C 3A）为探针，研究小干扰RNA（siRNA）-RNAi相关蛋白质相互作用。预期Br 7 C7 A和Br 3C 3A的溴取代基分别位于大沟和小沟中，并且当这些化学探针掺入siRNA中时在每个沟中充当空间位阻。使用含有这些化学探针的siRNA进行的全面研究表明，（i）乘客链5′端的Br 3C 3A（s）增强了它们的RNAi活性，（ii）RISC组装的方向由RISC的主要组分Argonaute 2与乘客链5′端附近小沟中的siRNA之间的相互作用决定。利用这些化学探针能够研究RNA和蛋白质之间的动态相互作用。
Elucidation of dynamic interactions between RNA and proteins is essential for understanding the biological processes regulated by RNA, such as RNA interference (RNAi). In this study, the logical chemical probes, comprising 7-bromo-7-deazaadenosine (Br7C7A) and 3-bromo-3-deazaadenosine (Br3C3A), to investigate small interfering RNA (siRNA)–RNAi related protein interactions, were developed. The bromo substituents of Br7C7A and Br3C3A are expected to be located in the major and the minor grooves, respectively, and to act as a steric hindrance in each groove when these chemical probes are incorporated into siRNAs. A comprehensive investigation using siRNAs containing these chemical probes revealed that (i) Br3C3A(s) at the 5′-end of the passenger strand enhanced their RNAi activity, and (ii) the direction of RISC assembly is determined by the interaction between Argonaute2, which is the main component of RISC, and siRNA in the minor groove near the 5′-end of the passenger strand. Utilization of these chemical probes enables the investigation of the dynamic interactions between RNA and proteins.