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首页> 外文期刊>Biochemistry >UNAMBIGUOUS STRUCTURE CHARACTERIZATION OF A DNA-RNA TRIPLE HELIX BY N-15- AND C-13-FILTERED NOESY SPECTROSCOPY
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UNAMBIGUOUS STRUCTURE CHARACTERIZATION OF A DNA-RNA TRIPLE HELIX BY N-15- AND C-13-FILTERED NOESY SPECTROSCOPY

机译：N-15和C-13过滤的鼻声波谱法对DNA-RNA三重螺旋的清晰结构表征

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DNA . DNA*RNA triple helices of the pyrimidine . purine*pyrimidine motif (where . indicates Watson-Crick pairing and * indicates Hoogsteen pairing) appear to be very stable, which has important implications for the development of novel antisense strategies. Here we present the first structural NMR studies on such a system, composed of a DNA hairpin with a homopurine-homopyrimidine stem sequence and a single-stranded RNA oligonucleotide containing exclusively pyrimidine residues. In these investigations an unlabeled DNA hairpin and a uniformly C-13/N-15-enriched RNA oligonucleotide were utilized in combination with X-edited H-1 NMR spectroscopy. Improved N-15 (omega(2)) filtered NOESY and C-13 (omega(1)) filtered NOESY are presented by which we were able to differentiate between intrastrand, i.e., DNA-DNA and RNA-RNA, and interstrand, i.e., DNA-RNA, NOE contacts. It is unambiguously established that the complex forms a right-handed triple helix, with the RNA strand situated in the major groove of the Watson-Crick stem of the hairpin. The interaction is stabilized by the formation of Hoogsteen-type base pairs between the RNA strand and the purine strand of the DNA. These strands run parallel to each other. The characterization of the DNA-RNA triple helix structure described here shows that this type of experiment forms a valuable instrument in the structure determination of bimolecular systems of nucleic acids.
机译：DNA。嘧啶的DNA * RNA三重螺旋。嘌呤*嘧啶基序（其中表示沃森-克里克配对，*表示Hoogsteen配对）似乎非常稳定，这对新型反义策略的开发具有重要意义。在这里，我们介绍了这种系统的首次结构NMR研究，该系统由具有高嘌呤-高嘧啶茎序列的DNA发夹和仅包含嘧啶残基的单链RNA寡核苷酸组成。在这些研究中，未标记的DNA发夹和均一的C-13 / N-15富集的RNA寡核苷酸与X编辑的H-1 NMR光谱结合使用。提出了改进的N-15（omega（2））过滤的NOESY和C-13（omega（1））过滤的NOESY，通过它们我们能够区分链内（即DNA-DNA和RNA-RNA）与链间（即， ，DNA-RNA，NOE接触。明确确定该复合物形成右手三螺旋，其RNA链位于发夹的Watson-Crick茎的主沟中。相互作用通过DNA的RNA链和嘌呤链之间Hoogsteen型碱基对的形成而稳定。这些股线彼此平行。此处描述的DNA-RNA三螺旋结构的特征表明，这种类型的实验在核酸双分子系统的结构测定中形成了一种有价值的工具。

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