NOVEL GUANOSINE REQUIREMENT FOR CATALYSIS BY THE HAIRPIN RIBOZYME
NOVEL GUANOSINE REQUIREMENT FOR CATALYSIS BY THE HAIRPIN RIBOZYME
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DOI：
10.1038/354320a0
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发表时间：
1991-11-28
期刊：
影响因子：
64.8
通讯作者：
BURKE, JM
中科院分区：
文献类型：
--
作者：
CHOWRIRA, BM;BERZALHERRANZ, A;BURKE, JM
人们对开发“设计核酶”以在体外和体内靶向破坏 RNA 非常感兴趣 1。具有新颖特异性的核酶工程需要详细了解核酶-底物相互作用，并对序列特异性进行严格评估。发夹核酶催化有效且可逆的位点特异性切割反应 2-4。我们使用诱变和体外选择策略来证明 RNA 切割和连接绝对需要紧邻切割连接位点 3' 的鸟苷。该 G 不是有效底物结合所必需的，相反，其 2-氨基是催化所需活性位点的重要组成部分。
THERE is much interest in the development of 'designer ribozymes' to target destruction of RNAs in vitro and in vivo 1. Engineering of ribozymes with novel specificities requires detailed knowledge of the ribozyme-substrate interaction, and a rigorous evaluation of sequence specificity. The hairpin ribozyme catalyses an efficient and reversible site-specific cleavage reaction 2-4. We have used mutagenesis and in vitro selection strategies to show that RNA cleavage and ligation has an absolute requirement for guanosine immediately 3' to the cleavage-ligation site. This G is not required for efficient substrate binding, rather, its 2-amino group is an essential component of the active site required for catalysis.