A novel mode of regulation of an RNA-cleaving DNAzyme by effectors that bind to both enzyme and substrate
A novel mode of regulation of an RNA-cleaving DNAzyme by effectors that bind to both enzyme and substrate
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DOI：
10.1006/jmbi.2001.4811
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发表时间：
2001-07-20
影响因子：
5.6
通讯作者：
Sen, D
中科院分区：
文献类型：
--
作者：
Wang, DY;Sen, D
我们描述了一种通过使用RNA和DNA效应物来控制RNA切割核酸酶（核酶和DNA酶）活性的新的和通用的策略。而在传统的异变构酶（包括核酶）的催化控制是通过结合效应分子的酶，在我们的策略DNA和RNA调节剂结合酶和底物。该系统的设计允许控制催化作用，即使在没有详细了解相关核酶或DNA酶的二级和三级结构的情况下。在这里，我们利用RNA和DNA形成分支的三向连接的能力来调节体外选择的“10-23”DNAzyme的RNA切割活性三个数量级。相对于其他不稳定和催化性差的酶-底物复合物，通过DNA或RNA“调节剂”诱导形成稳定和催化活性“三向”酶-底物-调节剂复合物的能力来进行控制。这种广泛调节的“三向”核酶/DNAzyme系统可能在体内发现效用，以引起一种RNA转录物的催化破坏，这取决于另一种基因转录物在其直接环境中的存在。(C)北京：科学出版社.
We describe a novel and general strategy for controlling the activity of RNA-cleaving nucleic acid enzymes (ribozymes and DNAzymes) via the use of RNA and DNA effecters. Whereas in conventional heteroallosteric enzymes (including ribozymes) control of catalysis is achieved by the binding of effector molecules to the enzyme, in our strategy DNA and RNA regulators bind to both the enzyme and the substrate. The design of this system permits the control of catalysis even in the absence of a detailed knowledge of the secondary and tertiary structure of the relevant ribozyme or DNAzyme. Here, we utilize the ability of RNA and DNA to form branched three-way junctions to regulate the RNA-cleaving activity of the in vitro selected "10-23" DNAzyme by three orders of magnitude. Control is exercised by the ability of a DNA or RNA "regulator" to induce formation of stable and catalytically competent "three-way" enzyme-substrate-regulator complexes, relative to otherwise unstable and catalytically poor enzyme-substrate complexes. Such expansively regulated "three-way" ribozyme/DNAzyme systems might find utility in vivo to bring about the catalyzed destruction of one RNA transcript contingent on the presence in its immediate environment of another gene transcript. (C) 2001 Academic Press.