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首页> 外文期刊>Nucleic Acids Research >UNEXPECTED ANISOTROPY IN SUBSTRATE CLEAVAGE RATES BY ASYMMETRIC HAMMERHEAD RIBOZYMES
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UNEXPECTED ANISOTROPY IN SUBSTRATE CLEAVAGE RATES BY ASYMMETRIC HAMMERHEAD RIBOZYMES

机译：不对称锤头状核酶在基质裂解率中的异乎寻常的各向异性

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摘要

RNA substrates which form relatively short helices I and III with hammerhead ribozymes are generally cleaved more rapidly than substrates which create longer binding helices, We speculated that for optimum cleavage rates, one of the helices needed to be relatively weak, To identify this helix, a series of ribozymes and substrates of varying lengths were made such that in the complex, helices I and III consisted of 5 and 10 bp respectively or vice versa, In two independent systems, substrates in the complexes with the shorter helix I and longer helix III were cleaved one to two orders of magnitude more rapidly than those in the complexes with the longer helix I and shorter helix III, Similar results were obtained whether the numbers of base pairs in helices I and III were limited either by the length of the hybridizing arms of the ribozyme or the length of the substrate, The phenomenon was observed for both all-RNA and DNA armed ribozymes, Thus, a relatively short helix I is required for fast cleavage rates in pre-formed hammerhead ribozyme-substrate complexes, When helix III has 10 bp, the optimum length for helix I is similar to 5 bp.
机译：通常，与锤头状核酶形成相对较短螺旋I和III的RNA底物的裂解速度要比产生较长结合螺旋的底物的裂解速度更快。我们推测，为了获得最佳裂解速率，其中一个螺旋必须相对较弱。制备了一系列不同长度的核酶和底物，使得在复合物中，螺旋I和III分别由5和10 bp组成，反之亦然。在两个独立的系统中，具有较短螺旋I和较长螺旋III的复合物中的底物是与具有较长螺旋I和较短螺旋III的复合物相比，其裂解速度要快一到两个数量级，无论螺旋I和III中碱基对的数目是否受杂交臂长度的限制，都获得了相似的结果核酶或底物的长度，这种现象在全RNA和DNA武装的核酶中都观察到，因此，需要相对较短的螺旋I在预先形成的锤头状核酶-底物复合物中，裂解速度很快，当螺旋III具有10 bp时，螺旋I的最佳长度接近5 bp。

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