Han H, Dervan P B
Beckman Institute, California Institute of Technology, Pasadena 91125.
Nucleic Acids Res. 1994 Jul 25;22(14):2837-44. doi: 10.1093/nar/22.14.2837.
Different helical conformations of DNA (D), RNA (R), and DNA.RNA (DR) hybrid double and triple helices have been detected using affinity cleavage analysis. Synthetic methods were developed to attach EDTA.Fe to a single nucleotide on RNA as well as DNA oligonucleotides. Cleavage patterns generated by a localized diffusible oxidant in the major groove on the pyrimidine strand of four purine.pyrimidine double helices consisting of all DNA, all RNA, and the corresponding hybrids reveal that the relative cleavage intensity shifts to the 5' end of the purine strand increasingly in the order: DD < DR < RD < RR. These results are consistent with models derived from structural studies. In six pyrimidine.purine.pyrimidine triple helices, the altered cleavage patterns of the Watson-Crick pyrimidine strands reveal at least two conformational families: (i) D + DD, R + DD, D + DR, and R + DR and (ii) R + RD and R + RR.
利用亲和裂解分析检测到了DNA(D)、RNA(R)以及DNA-RNA(DR)杂交双链和三链螺旋的不同螺旋构象。已开发出合成方法,将乙二胺四乙酸铁(EDTA.Fe)连接到RNA以及DNA寡核苷酸的单个核苷酸上。由一种局部可扩散氧化剂在四条嘌呤-嘧啶双链螺旋(由全DNA、全RNA以及相应杂交体组成)嘧啶链大沟中产生的裂解模式表明,相对裂解强度按以下顺序越来越多地向嘌呤链的5'端转移:DD < DR < RD < RR。这些结果与结构研究得出的模型一致。在六个嘧啶-嘌呤-嘧啶三链螺旋中,沃森-克里克嘧啶链裂解模式的改变揭示了至少两个构象家族:(i)D + DD、R + DD、D + DR和R + DR,以及(ii)R + RD和R + RR。