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首页> 外文期刊>Biochemistry >Nickel- and cobalt-dependent reagents identify structural features of RNA that are not detected by dimethyl sulfate or RNase Tl
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Nickel- and cobalt-dependent reagents identify structural features of RNA that are not detected by dimethyl sulfate or RNase Tl

机译：镍和钴依赖性试剂可识别硫酸二甲酯或RNase T1未检测到的RNA结构特征

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摘要

Ribosomal 5S RNA presents a particular challenge to structural investigations since this polynucleotide is too large for complete NMR characterization but lacks significant tertiary structure to modulate, for example, diagnostic alkylation of guanine N7 by dimethyl sulfate. Nickel-and cobalt-dependent reagents that are sensitive to the N7 and aromatic face of guanine have now been applied to 5S rRNA (Xenopus lavis) and provide structural information that was not previously available from traditional chemical or enzymatic probes. Although G75 had repeatedly demonstrated an average reactivity with dimethyl sulfate and minimal reactivity with RNase T1, this residue was the major target of both metal-dependent reagents. Such reactivity provides crucial support for a structural model of loop E identified by prior physical, but not chemical, methods. Similarly, the tetraloop structure of loop D was more accurately reflected by the reactivity of G87 and G89 in the presence of the nickel reagent rather than in the presence of RNase T1. In addition, nickel-dependent modification of guanine residues surrounding the three-helix junction of loop A suggests an organization that is less compact than previously considered. [References: 66]
机译：核糖体5S RNA对结构研究提出了特殊的挑战，因为该多核苷酸太大，无法进行完整的NMR表征，但缺少显着的三级结构，无法调节例如硫酸二甲酯对鸟嘌呤N7的诊断烷基化。对鸟嘌呤N7和芳香面敏感的镍和钴依赖性试剂现已应用于5S rRNA（非洲爪蟾），并提供了传统化学或酶探针无法获得的结构信息。尽管G75反复证明了与硫酸二甲酯的平均反应性和与RNase T1的最小反应性，但该残基是两种金属依赖性试剂的主要目标。这种反应性为通过现有物理方法而非化学方法鉴定的环E的结构模型提供了关键的支持。类似地，在镍试剂存在下而不是在RNase T1存在下，G87和G89的反应性更准确地反映了环D的四环结构。另外，围绕环A的三螺旋连接点的鸟嘌呤残基的镍依赖性修饰表明该结构比先前考虑的更不紧密。 [参考：66]

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