Restoration of Ribozyme Tertiary Contact and Function by Using a Molecular Glue for RNA
Restoration of Ribozyme Tertiary Contact and Function by Using a Molecular Glue for RNA
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DOI：
10.1002/anie.201709041
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发表时间：
2018-01-08
影响因子：
16.6
通讯作者：
Nakatani, Kazuhiko
中科院分区：
文献类型：
--
作者：
Dohno, Chikara;Kimura, Maki;Nakatani, Kazuhiko
一些RNA类需要折叠成适当的高阶结构才能发挥它们的功能。锤头状核酶(HHR)需要一个由三级相互作用稳定的折叠构象才能发挥全部活性。合成的RNA结合配体N-氨基甲酸四聚体Z-二苯乙烯连接物(Z-NCTS)可以激活非活性的HHR。Z-NCTS的结合可以诱导活性折叠结构的形成，从而恢复核酶活性，其中Z-NCTS充当分子粘合剂，使两个分离的RNA环相互接触。接下来，我们利用插入到3‘非翻译区的HHR序列作为顺式作用元件设计了一个Z-NCTS响应型遗传开关。我们证明，设计合理的核酶开关能够通过Z-NCTS调节基因表达，并在哺乳动物细胞中发挥作用。
Some RNA classes require folding into the proper higher-order structures to exert their functions. Hammerhead ribozyme (HHR) requires a folding conformation stabilized by tertiary interaction for full activity. A rationally engineered HHR was developed that was inactive, but could be activated by a synthetic RNA-binding ligand, naphthyridine carbamate tetramer with Z-stilbene linker (Z-NCTS). Binding of Z-NCTS could induce the formation of an active folding structure and thereby restore ribozyme activity, where Z-NCTS acts as a molecular glue to bring two isolated RNA loops into contact with each other. Next, we designed a Z-NCTS-responsive genetic switch using the HHR sequence inserted into the 3' untranslated region as a cis-acting element. We demonstrated that the rationally designed ribozyme switch enabled regulation of gene expression by Z-NCTS and was functional in mammalian cells.