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在通用支架上定制RNA模块化单元:一种具有催化单元的模块化核酶,用于β-烟酰胺单核苷酸激活的RNA连接。

Tailoring RNA modular units on a common scaffold: a modular ribozyme with a catalytic unit for beta-nicotinamide mononucleotide-activated RNA ligation.

作者信息

Fujita Yuki, Furuta Hiroyuki, Ikawa Yoshiya

机构信息

Department of Chemistry and Biochemistry, Graduate School of Engineering, Kyushu University, Fukuoka 819-0395, Japan.

出版信息

RNA. 2009 May;15(5):877-88. doi: 10.1261/rna.1461309. Epub 2009 Mar 23.

Abstract

A novel ribozyme that accelerates the ligation of beta-nicotinamide mononucleotide (beta-NMN)-activated RNA fragments was isolated and characterized. This artificial ligase ribozyme (YFL ribozyme) was isolated by a "design and selection" strategy, in which a modular catalytic unit was generated on a rationally designed modular scaffold RNA. Biochemical analyses of the YFL ribozyme revealed that it catalyzes RNA ligation in a template-dependent manner, and its activity is highly dependent on its architecture, which consists of a modular scaffold and a catalytic unit. As the design and selection strategy was used for generation of DSL ribozyme, isolation of the YFL ribozyme indicated the versatility of this strategy for generation of functional RNAs with modular architectures. The catalytic unit of the YFL ribozyme accepts not only beta-NMN but also inorganic pyrophosphate and adenosine monophosphate as leaving groups for RNA ligation. This versatility of the YFL ribozyme provides novel insight into the possible roles of beta-NMN (or NADH) in the RNA world.

摘要

一种能够加速β -烟酰胺单核苷酸(β -NMN)激活的RNA片段连接的新型核酶被分离并鉴定。这种人工连接酶核酶(YFL核酶)是通过“设计与筛选”策略分离得到的,即在合理设计的模块化支架RNA上构建模块化催化单元。对YFL核酶的生化分析表明,它以模板依赖的方式催化RNA连接,其活性高度依赖于由模块化支架和催化单元组成的结构。由于“设计与筛选”策略用于生成DSL核酶,YFL核酶的分离表明该策略在生成具有模块化结构的功能性RNA方面具有通用性。YFL核酶的催化单元不仅接受β -NMN作为RNA连接的离去基团,还接受无机焦磷酸和单磷酸腺苷。YFL核酶这种通用性为β -NMN(或NADH)在RNA世界中可能扮演的角色提供了新的见解。

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