Suh W C, Ross W, Record M T
Department of Chemistry, University of Wisconsin, Madison 53706.
Science. 1993 Jan 15;259(5093):358-61. doi: 10.1126/science.8420002.
Potassium permanganate (KMnO4) footprinting in the absence and presence of magnesium (Mg2+) at the lambda PR promoter identified two different open complexes with Escherichia coli E sigma 70 RNA polymerase (designated RPo1 and RPo2). The single-stranded region in RPo1 (formed in the absence of Mg2+) was at most 12 bases long, whereas that in RPo2 (formed in the presence of Mg2+) spanned at least 14 bases. Only in RPo2 did the single-stranded region extend to the start point of transcription (+1, +2). These results provide a structural basis for the requirement for uptake of Mg2+ in the formation of RPo2 from RPo1, as deduced from kinetic studies at this promoter.
在有无镁离子(Mg2+)存在的情况下,针对λPR启动子进行的高锰酸钾(KMnO4)足迹分析,鉴定出了两种不同的与大肠杆菌E σ70 RNA聚合酶形成的开放复合物(分别命名为RPo1和RPo2)。RPo1(在无Mg2+存在时形成)中的单链区域最长为12个碱基,而RPo2(在有Mg2+存在时形成)中的单链区域至少跨越14个碱基。只有在RPo2中,单链区域才延伸至转录起始点(+1,+2)。这些结果为从动力学研究中推断出的在该启动子处从RPo1形成RPo2时对Mg2+摄取的需求提供了结构基础。