MAPPING OF PSORALEN CROSS-LINKED NUCLEOTIDES IN RNA
MAPPING OF PSORALEN CROSS-LINKED NUCLEOTIDES IN RNA
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DOI：
10.1093/nar/12.7.3405
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发表时间：
1984-01-01
影响因子：
14.9
通讯作者：
KUMAR, A
中科院分区：
文献类型：
--
作者：
GARRETTWHEELER, E;LOCKARD, RE;KUMAR, A
描述了一种使用交联剂 4'-(羟甲基)-4,5',8-三甲基补骨脂素 (HMT) 绘制 RNA 分子内 bsss 配对区域和高阶结构的方法。将此方法应用于酵母 tRNAPhe，我们专门鉴定了 C11X C25 之间的 D 茎中的 U6X U68 之间的受体茎内以及 U50X C63 和 U52X C63 之间的 Tψ 茎中的交联。我们还发现了 U8X C48 之间的独特交联，由于 U8:A14 和 C48:G15 之间的三级相互作用，U8X C48 是核心区域的反式嘧啶。在每种情况下，通过使用嘌呤特异性 U2 核糖核酸酶和胞苷特异性 CL3 核糖核酸酶（在光反转嘧啶后会异常裂解），推导出精确的交联点。绘制精确交联点图谱的能力对于识别其他 RNA 分子三级结构中非常接近的核苷酸来说应该是无价的。
A method is described for using the cross-linking reagent 4′-(hydroxy-methyl)-4,5′,8-trimethylpsoralen (HMT) to map bsss paired regions and higher-order structure within RNA molecules. Applying this method to yeast tRNAPhe, we have specifically identified cross-links within the acceptor stem between U6X U68in the D-stem between C11X C25and in the Tψ-stem between U50X C63and U52X C63. We have also identified a unique cross-link between U8X C48which are trans pyrimidines in the core region due to tertiary interactions between U8:A14and C48:G15The precise point of cross-linking was deduced in every case by using purine-specific U2ribo-nuclease along with cytidine-specific CL3 ribonuclease which will anomalously cleave after photoreversed pyrimidines. The ability to map the precise point of cross-linking should prove invaluable in identifying nucleotides in close proximity within the tertiary structure of other RNA molecules.