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首页> 外文期刊>Biophysical Chemistry: An International Journal Devoted to the Physical Chemistry of Biological Phenomena >Probing the structure of multi-stranded guanine-rich DNA complexes by Raman spectroscopy and enzymatic degradation.
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Probing the structure of multi-stranded guanine-rich DNA complexes by Raman spectroscopy and enzymatic degradation.

机译：通过拉曼光谱和酶促降解探索多链富含鸟嘌呤的DNA复合物的结构。

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摘要

The multi-stranded DNA complexes formed by the oligonucleotides d(T15G4T2G4), Tel, and d(T15G15), TG, were examined by nuclease digestion and Raman spectroscopy. Both Tel and TG can aggregate to form structures consisting of multiple, parallel-oriented DNA strands with two independent structural domains. Overall, the structures of the TG and Tel aggregates appear similar. According to the Raman data, the majority of bases are in C2'-endo/anti conformation. The interaction of guanines at the 3'-ends in both complexes stabilizes the complexes and protects them from degradation by exonuclease III. The 5'-extensions remain single-stranded and the thymines are accessible to single-strand-specific nuclease digestion. The extent of enzymatic cleavage at the junction at the 5' end of the 15 thymines implies a conformational change between this part of the molecule and the guanine-rich region. The differential enzymatic sensitivity of the complexes suggests there are variations in backbone conformations between TG and Tel aggregates. TG aggregates were more resistant to digestion by DNase I, Mung Bean nuclease, and S1 nuclease than Tel complexes. It is proposed that the lower DNase I sensitivity may be partly due to the more stable backbone exhibited by TG than Tel complexes. Structural uniformity along the guanine core of TG is suggested, as there is no indication of structural discontinuities or protected sites in the guanine-rich regions of TG aggregates. The lower extent of digestion by Mung Bean nuclease at the 3' end implies that these bases are inaccessible to the enzyme. This suggests that there is minimal fraying at the ends, which is consistent with the extreme thermal stability of the TG aggregates.
机译：通过核酸酶消化和拉曼光谱检查了由寡核苷酸d（T15G4T2G4）Tel和d（T15G15）TG形成的多链DNA复合物。 Tel和TG均可聚集形成由具有两个独立结构域的多个平行定向的DNA链组成的结构。总体而言，TG和Tel聚集体的结构看起来相似。根据拉曼数据，大多数碱基处于C2'-内/反构象。两种复合物中3'端鸟嘌呤的相互作用使复合物稳定并保护它们免受核酸外切酶III的降解。 5'-延伸区保持单链，胸腺嘧啶可用于单链特异性核酸酶消化。在15个胸腺嘧啶的5'末端连接处的酶促裂解程度暗示了该分子的这一部分与鸟嘌呤富集区之间的构象变化。复合物的不同酶敏感性表明，TG和Tel聚集体之间的骨架构象存在差异。 TG聚集体比DN复合体对DNase I，绿豆核酸酶和S1核酸酶的消化更有抵抗力。提出较低的DNase I敏感性可能部分归因于TG显示的骨架比Tel复合物更稳定。建议沿TG的鸟嘌呤核结构均匀，因为在TG聚集体的鸟嘌呤丰富的区域中没有结构不连续或受保护的部位的迹象。绿豆核酸酶在3'端的消化程度较低，表明这些碱基无法进入该酶。这表明在末端的磨损最小，这与TG聚集体的极高热稳定性相一致。

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