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首页> 外文期刊>Journal of Molecular Biology >Hairpin ribozymes with four-way helical junctions mediate intracellular RNA ligation
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Hairpin ribozymes with four-way helical junctions mediate intracellular RNA ligation

机译：具有四向螺旋连接的发夹状核酶介导细胞内RNA连接

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Virtually all RNA-mediated reactions require transitions among alternative RNA conformations. The complexity of biological reactions can obscure specific conformational changes in vivo and important features of the intracellular environment are difficult to reproduce in vitro. However, simple RNA self-cleavage and ligation reactions offer a unique opportunity to measure the kinetics and equilibria of specific RNA conformational transitions directly in living cells. Hairpin ribozymes that incorporate the natural four-way helical junction self-cleave rapidly in vivo, but only when cleavage products dissociate rapidly. Cleavage rates fall when cleavage products remain bound in stable base-paired helices, providing evidence that bound products undergo re-ligation. These results provide the first detailed kinetic description of an intracellular ribozyme reaction that includes cleavage, ligation and product dissociation rates. Kinetic and equilibrium parameters measured in vivo correspond well, but not perfectly, with values measured for the same reactions in vitro under conditions that approximate an intracellular ionic environment. (C) 2001 Academic Press. [References: 35]
机译：实际上，所有RNA介导的反应都需要在其他RNA构象之间进行转换。生物反应的复杂性会掩盖体内特定的构象变化，而细胞内环境的重要特征很难在体外复制。然而，简单的RNA自切割和连接反应提供了一个独特的机会，可以直接在活细胞中测量特定RNA构象转变的动力学和平衡。掺入天然四向螺旋连接的发夹状核酶会在体内快速自我裂解，但仅在裂解产物迅速解离时才如此。当裂解产物保持结合在稳定的碱基配对的螺旋中时，裂解率下降，这提供了结合产物经历重新连接的证据。这些结果提供了细胞内核酶反应的第一个详细的动力学描述，该反应包括裂解，连接和产物解离速率。在体内测得的动力学和平衡参数与在近似细胞内离子环境的条件下在体外对相同反应测得的值相吻合，但并不完美。 （C）2001学术出版社。 [参考：35]

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