Lewis J B, Brass L F, Doty P
Biochemistry. 1975 Jul 15;14(14):3164-71. doi: 10.1021/bi00685a020.
The binding of oligonucleotides to synthetic polynucleotides has been studied as a control for investigations of the binding of oligonucleotides to natural RNA molecules. Only combinations that involved A-U, G-C, and G-G pairs were found to be significantly stable under the experimental conditions used here. The stability of the oligomerpolymer pairing increased with the length of the region paired and with its G + C content. Further, some different sequence isomers of the same G + C content exhibited quite different binding constants. This variability is consistent with certain sequence differences in the double-strand stacking interactions stabilizing the oligomer-polymer association. Oligomer binding was also shown to depend upon the identity of the polymer residues neighboring the binding site, indicating the effect upon oligomer binding of small changes in the single-strand conformation of the binding site. These observations validate the criteria that allow one to decide if an observed association constant of an oligomer to an RNA molecule reflects a complete complementarity between the two or not. This improves the basis for using oligonucleotide binding constants to RNA of known sequences to map secondary structure.
已对寡核苷酸与合成多核苷酸的结合进行了研究,以此作为寡核苷酸与天然RNA分子结合研究的对照。在此处使用的实验条件下,仅发现涉及A-U、G-C和G-G碱基对的组合具有显著稳定性。寡聚物-聚合物配对的稳定性随配对区域的长度及其G+C含量的增加而增加。此外,相同G+C含量的一些不同序列异构体表现出截然不同的结合常数。这种变异性与稳定寡聚物-聚合物缔合的双链堆积相互作用中的某些序列差异一致。寡聚物结合还显示取决于结合位点相邻的聚合物残基的特性,这表明结合位点单链构象的微小变化对寡聚物结合的影响。这些观察结果验证了一些标准,这些标准可让人判断观察到的寡聚物与RNA分子的缔合常数是否反映了两者之间的完全互补性。这改善了使用已知序列的RNA的寡核苷酸结合常数来绘制二级结构的基础。