Mishima Y, Suda T, Kominami R
Department of Biochemistry, Niigata University School of Medicine.
J Biochem. 1996 Apr;119(4):805-10. doi: 10.1093/oxfordjournals.jbchem.a021311.
Incubation of the 162-bp duplex containing d(GGA:TCC)11 repeats with d(GGA)11 and d(GGT)11 oligonucleotides, but not with d(TCC)11, gave a mobility-shifted band in polyacrylamide gel electrophoresis, suggestive of the triplex formation. The two conformers, however, showed difference in binding affinity and structure. The complex formed with d(GGT)11 exhibited patterns protected from DNase I digestion and from modifications with osmium tetroxide and dimethyl sulfate. The patterns were conformed to a proposed triplex model. In contrast, the complex with d(GGA)11 was sensitive to DNase I and to osmium tetroxide, which is reactive to pyrimidine bases of single-stranded DNA. These results suggest that the d(GGA:TCC)11 and d(GGA)11 triple-stranded complex forms a D-loop-like structure. By considering the ability of d(GGA)11 oligonucleotides to form a parallel homoduplex, the structure may be one in which the GGA-strand of the duplex pairs with d(GGA)11 in parallel, with the TCC-strand being looped out.
将含有d(GGA:TCC)11重复序列的162碱基对双链体与d(GGA)11和d(GGT)11寡核苷酸一起温育,而不是与d(TCC)11一起温育,在聚丙烯酰胺凝胶电泳中产生了一条迁移率改变的条带,提示形成了三链体。然而,这两种构象在结合亲和力和结构上存在差异。与d(GGT)11形成的复合物表现出对DNase I消化以及四氧化锇和硫酸二甲酯修饰的抗性模式。这些模式符合提出的三链体模型。相比之下,与d(GGA)11形成的复合物对DNase I和四氧化锇敏感,四氧化锇对单链DNA的嘧啶碱基有反应。这些结果表明,d(GGA:TCC)11和d(GGA)11三链体复合物形成了一种D环样结构。考虑到d(GGA)11寡核苷酸形成平行同源双链体的能力,该结构可能是双链体的GGA链与d(GGA)11平行配对,而TCC链形成环的结构。