In vitro selection of an archaeal RNase P RNA mimics natural variation.
In vitro selection of an archaeal RNase P RNA mimics natural variation.
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DOI：
10.1155/2004/903283
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发表时间：
2004-10
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影响因子：
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通讯作者：
Daniel Williams;James W. Brown
中科院分区：
文献类型：
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作者：
Daniel Williams;James W. Brown
古细菌和细菌的RNase P rna在序列和二级结构上是相似的，但在缺乏蛋白质的情况下，古细菌rna的活性要低得多，需要极端的离子条件才能活性。为了评估古细菌RNA的活性如何容易地通过序列的微小变化来提高，我们使用体外选择来产生甲酸甲烷杆菌RNase P RNA的变体：芽孢杆菌subtilus pre-tRNA（Asp）自裂偶联RNA。功能变异是由一系列突变产生的，这些突变主要与该RNA的自然变异一致。由选择产生的变异具有与野生型相当的裂解率；没有获得裂解率提高或离子需求量降低的变体。这说明细菌和古生菌的RNase P RNA得到了全局优化，两种RNase P RNA存在较大生化差异的基础是分布在分子内的。
Archaeal and bacterial RNase P RNAs are similar in sequence and secondary structure, but in the absence of protein, the archaeal RNAs are much less active and require extreme ionic conditions for activity. To assess how readily the activity of the archaeal RNA alone could be improved by small changes in sequence, in vitro selection was used to generate variants of a Methanobacterium formicicum RNase P RNA: Bacillus subtilus pre-tRNA(Asp) self-cleaving conjugate RNA. Functional variants were generated with a spectrum of mutations that were predominately consistent with natural variation in this RNA. Variants generated from the selection had cleavage rates comparable to that of wild type; variants with improved cleavage rates or lower ionic requirements were not obtained. This suggests that the RNase P RNAs of Bacteria and Archaea are globally optimized and the basis for the large biochemical differences between these two types of RNase P RNA is distributed in the molecule.