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FORMATION OF A PARALLEL-STRANDED DNA HOMODUPLEX BY D(GGA) REPEAT OLIGONUCLEOTIDES

机译:D(GGA)重复寡核苷酸形成的平行链DNA同源多肽

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摘要

The GGA9-H molecules consisting of a double helical stretch followed by a single-stranded 3'-terminal overhang of nine GGA sequence repeats exhibited a gel mobility-shifted band in a concentration-dependent manner, suggestive of the intermolecular complex formation. The position of the shifted band in a gel was almost identical to that of the Y-shaped dimer marker of the same molecular weight that had the two double-helices at one side, This suggests that GGA9-H dimerizes in a parallel orientation without the formation of four-stranded hairpin structure, Since the GGA9-H homoduplex was stably formed at pH 4, 7 and 9, the formation does not require protonation or deprotonation of the N1 position of adenines, Neither does it require the N7 group of guanines responsible for Hoogsteen base pairing from the methylation interference and modification studies, Modification of the N7 group of guanines with dimethyl sulfate (DMS) did not inhibit the association and also the N7 group in the homoduplex was not protected from DMS, On the other hand, the GAA9-H having the G to A base substitution did not show such an association with either GGA9-H or GAA9-H. These results suggest that the homoduplex formation may be due to G G base pairing through non-Hoogsteen hydrogen bonds.
机译:由九个GGA序列重复序列的双螺旋拉伸和单链3'-末端突出组成的GGA9-H分子以浓度依赖的方式显示出凝胶迁移率移动的条带,表明分子间复合物的形成。凝胶中移位带的位置几乎与分子量相同的Y形二聚体标记的位置相同,该标记在一侧具有两个双螺旋,这表明GGA9-H在平行方向上二聚,而没有形成四链发夹结构,由于GGA9-H同双链体在pH 4、7和9稳定形成,因此不需要腺嘌呤N1位置的质子化或去质子化,也不需要负责鸟嘌呤的N7基团对于来自甲基化干扰和修饰研究的Hoogsteen碱基配对,鸟嘌呤的N7基团被硫酸二甲酯修饰(DMS)不会抑制该缔合,同双链体中的N7基团也不受DMS保护,另一方面,具有G至A碱基取代的GAA9-H与GGA9-H或GAA9-H均未显示出这种关联。这些结果表明同源双链体的形成可能是由于通过非Hoogsteen氢键的G G碱基配对所致。

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