正交安装到接口上，组装四膜虫组 I 核酶的两个模块域。

Installation of orthogonality to the interface that assembles two modular domains in the Tetrahymena group I ribozyme.

机构信息

Department of Chemistry and Biochemistry, Graduate School of Engineering, Kyushu University, Moto-oka 744, Nishi-ku, Fukuoka 819-0395, Japan.

Department of Chemistry and Biochemistry, Graduate School of Engineering, Kyushu University, Moto-oka 744, Nishi-ku, Fukuoka 819-0395, Japan; International Research Center for Molecular Systems, Kyushu University, Moto-oka 744, Nishi-ku, Fukuoka 819-0395, Japan.

出版信息

J Biosci Bioeng. 2014 Apr;117(4):407-12. doi: 10.1016/j.jbiosc.2013.10.008. Epub 2013 Nov 8.

DOI:10.1016/j.jbiosc.2013.10.008

PMID:24216461

Abstract

Two modular elements (P5abc and ΔP5) in the Tetrahymena group I ribozyme can be separated physically to generate a two-piece ribozyme derivative consisting of a separately prepared P5abc (P5 RNA) and the rest of the intron (ΔP5 RNA). Molecular recognition in the interface assembling P5 RNA and ΔP5 RNA is strong and specific, and the catalytic ability of the two-piece ribozyme is comparable to that of the parent unimolecular ribozyme. We designed alternative P14 (L5c-L2) interacting modules participating in the assembly of P5 and ΔP5 and investigated their ability in the context of complex formation of the two-piece ribozyme and in vivo splicing of the unimolecular intron ribozyme. Combined use of alternative P14 and L5b-P6 interacting modules provided robust orthogonality to the P5/ΔP5 assembly interface of the bimolecular complex.

摘要

两个 Tetrahymena 组 I 核酶的模块化元件（P5abc 和 ΔP5）可以在物理上分离，生成由单独制备的 P5abc（P5 RNA）和内含子其余部分（ΔP5 RNA）组成的两段式核酶衍生物。在组装 P5 RNA 和 ΔP5 RNA 的界面处的分子识别是强特异性的，两段式核酶的催化能力与母本单分子核酶相当。我们设计了替代的 P14（L5c-L2）相互作用模块，参与 P5 和 ΔP5 的组装，并在两段式核酶的复杂形成和单分子内含子核酶的体内剪接的背景下研究了它们的能力。替代的 P14 和 L5b-P6 相互作用模块的组合使用为双分子复合物的 P5/ΔP5 组装界面提供了强大的正交性。