Wang Y H, Howard M T, Griffith J D
Lineberger Cancer Research Center, University of North Carolina, Chapel Hill 27514.
Biochemistry. 1991 Jun 4;30(22):5443-9. doi: 10.1021/bi00236a017.
Tracts of four to six adenines phased with the DNA helix produce a sequence-directed bending of the helix axis. Here, using gel electrophoresis and electron microscopy (EM), we have asked whether a similar motif will induce bending in a duplex RNA helix. Single-stranded RNAs were transcribed either from short synthetic DNA templates or from Crithidia fasciculata kinetoplast bent DNA, and the complementary single-stranded RNAs were annealed to produce duplex RNA molecules containing blocks of four to six adenines. Electrophoresis on polyacrylamide gels revealed no retardation of the RNAs containing phased blocks of adenines relative to duplex RNAs lacking such blocks. Examination by EM showed most of the molecules to be straight or only slightly bent. Thus, in contrast to DNA duplexes, phased adenine tracts do not induce sequence-directed bending in double-stranded RNA. Analysis of the distribution of molecule shapes for the highly bent C. fasciculata DNA showed that the adenine blocks do not act cooperatively to induce DNA bending and that the molecules must equilibrate between a spectrum of bent shapes.
与DNA螺旋相匹配的四到六个腺嘌呤序列会使螺旋轴产生序列导向的弯曲。在此,我们通过凝胶电泳和电子显微镜(EM)研究了类似的基序是否会在双链RNA螺旋中诱导弯曲。单链RNA是从短的合成DNA模板或从克氏锥虫动质体弯曲DNA转录而来的,互补的单链RNA退火后产生含有四到六个腺嘌呤块的双链RNA分子。聚丙烯酰胺凝胶电泳显示,相对于缺乏此类块的双链RNA,含有腺嘌呤相控块的RNA没有延迟迁移。电子显微镜检查显示,大多数分子是直的或只是轻微弯曲。因此,与DNA双链体不同,相控腺嘌呤序列不会在双链RNA中诱导序列导向的弯曲。对高度弯曲的克氏锥虫DNA分子形状分布的分析表明,腺嘌呤块不会协同作用来诱导DNA弯曲,并且分子必须在一系列弯曲形状之间达到平衡。