Sequence-dependent structural variations of hammerhead RNA enzymes.
Sequence-dependent structural variations of hammerhead RNA enzymes.
复制标题
锤头状 RNA 酶的序列依赖性结构变异。
DOI：
10.1093/nar/18.5.1103
复制
发表时间：
1990
影响因子：
14.9
通讯作者：
Pardi,A
中科院分区：
文献类型：
--
作者：
Heus,HA;Uhlenbeck,OC;Pardi,A
锤头状RNA自切割结构域的体内催化活性的发现导致了一类新的序列特异性RNA内切酶的发展。用T7聚合酶体外转录法合成了两种这样的核酶，并用光谱、核磁共振和非变性凝胶电泳研究了它们的结构。这些数据显示在两种RNA酶中存在由双螺旋茎和四核苷环组成的稳定发夹。在两种RNA酶中也观察到具有不同稳定性的额外结构。这两种RNA酶切割互补RNA底物的半衰期先前已显示相差50倍。这里提出的数据表明，这种速率差异可能是由于在RNA酶中形成催化失活构象，这与酶-底物复合物的形成相互干扰。
The discovery ofin vivocatalytic activity for the hammerhead RNA self-cleaving domain has led to the development of a new class of sequence-specific RNA endonucleases. Two such ribozymes have been synthesized usingin vitrotranscription with T7 polymerase and their structures have been studied by optical spectroscopy, nuclear magnetic resonance and nondenaturing gel electrophoresis. These data show the presence of a stable hairpin consisting of a double helical stem and a tetranucleotlde loop in both RNA enzymes. Additional structure, with different stabilities, Is also observed in both RNA enzymes. The half-lives for cleavage of the complementary RNA substrates by these two RNA enzymes have been previously shown to differ by a factor of 50. The data presented here suggest that this rate difference may be a result of the formation of catalytically inactive conformations In the RNA enzyme which Interfere with formation of the enzyme-substrate complex.