Lead-catalyzed cleavage of ribonuclease P RNA as a probe for integrity of tertiary structure.
Lead-catalyzed cleavage of ribonuclease P RNA as a probe for integrity of tertiary structure.
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铅催化核糖核酸酶 P RNA 的裂解作为三级结构完整性的探针。
DOI：
10.1093/nar/21.25.5916
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发表时间：
1993
影响因子：
14.9
通讯作者：
Pace,NR
中科院分区：
文献类型：
--
作者：
Zito,K;Hüttenhofer,A;Pace,NR
Pb2+催化的RNA裂解是研究三级结构的有用探针。在本研究中，鉴定了来自三种系统发育完全不同的生物--大肠杆菌、花色杆菌和枯草杆菌的核糖核酸酶P RNA的Pb2+裂解模式，以及被缺失改变的E.coliRNase P RNA的Pb2+裂解模式。每个天然RNA在核心结构中的几个位置经历切割，这是所有细菌RNaseP RNA共同的。所有的切割都发生在RNA二级结构模型的非配对区域，在可能参与三级相互作用的区域。在所有天然RNA中，无论序列变化如何，都在同源位置存在两个切割位点，这表明它们具有共同的三级结构特征。4个缺失突变体的Pb2+裂解位点与天然模式不同，表明突变体RNA的三级结构发生了变化。这一结论与先前描述的突变RNA的性质一致。因此，Pb2+裂解试验是揭示RNaseP RNA三级结构变化的有用探针。未知结构(在图1中显示为未配对)，可能参与三级相互作用。在所有天然RNA中，无论序列变化如何，都在同源位置存在两个切割位点，这表明它们具有共同的三级结构特征。在E.coliRNase P RNA的4个缺失突变体中，Pb2+的切割位点与天然模式不同，表明突变的RNA的三级结构发生了变化。这一结论与先前描述的突变RNA的性质一致。因此，Pb2+裂解试验是揭示RNaseP RNA三级结构变化的有用探针。
Pb2+-catalyzed cleavage of RNA has been shown previously to be a useful probe for tertiary structure. In the present study, Pb2+cleavage patterns were identified for ribonuclease P RNAs from three phylogenetically disparate organisms,Escherichia coli, Chromatfum vinosum, Bacillus subtilis,and forE.coliRNase P RNAs that had been altered by deletions. Each of the native RNAs undergoes cleavage at several sites in the core structure that is common to all bacterial RNase P RNAs. All the cleavages occur in non-paired regions of the secondary structure models of the RNAs, in regions likely to be involved in tertiary interactions. Two cleavage sites occur at homologous positions in all the native RNAs, regardless of sequence variation, suggesting common tertiary structural features. The Pb2+cleavage sites in four deletion mutants of E.coli RNase P RNA differed from the native pattern, indicating alterations in the tertiary structures of the mutant RNAs. This conclusion is consistent with previously characterized properties of the mutant RNAs. The Pb2+cleavage assay is thus a useful probe to reveal alteration of tertiary structure in RNase P RNA. of yet-unknown structure (shown in Fig. 1 as unpaired), which are likely involved in tertiary interactions. Two cleavage sites occur at homologous positions in all the native RNAs, regardless of sequence variation, suggesting common tertiary structural features. The Pb2+cleavage sites in four deletion mutants ofE.coliRNase P RNA differed from the native pattern, indicating alterations in the tertiary structures of the mutant RNAs. This conclusion is consistent with previously characterized properties of the mutant RNAs. The Pb2+cleavage assay is thus a useful probe to reveal alteration of tertiary structure in RNase P RNA.