Meier N, Wagner R
Eur J Biochem. 1985 Jan 2;146(1):83-7. doi: 10.1111/j.1432-1033.1985.tb08622.x.
70S ribosomes and 30S and 50S ribosomal subunits from Escherichia coli were modified under non-denaturing conditions with the chemical reagent dimethylsulfate. The ribosomal 23S and 16S RNAs were isolated after the reaction and the last 200 nucleotides from the 3' ends were analyzed for differences in the chemical modification. A number of accessibility changes could be detected for 23S and 16S RNA when 70S ribosomes as opposed to the isolated subunits were modified. In addition to a number of sites which were protected from modification several guanosines showed enhanced reactivities, indicating conformational changes in the ribosomal RNA structures when 30S and 50S subunits associate to a 70S particle. Most of the accessibility changes can be localized in double-helical regions within the secondary structures of the two RNAs. The results confirm the importance of the ribosomal RNAs for ribosomal functions and help to define the RNA domains which constitute the subunit interface of E. coli ribosomes.
在非变性条件下,用化学试剂硫酸二甲酯对来自大肠杆菌的70S核糖体以及30S和50S核糖体亚基进行修饰。反应后分离出核糖体23S和16S RNA,并对3'端的最后200个核苷酸进行化学修饰差异分析。当修饰70S核糖体而非分离的亚基时,可检测到23S和16S RNA的许多可及性变化。除了一些免受修饰的位点外,几个鸟苷显示出增强的反应活性,表明当30S和50S亚基结合形成70S颗粒时,核糖体RNA结构发生了构象变化。大多数可及性变化可定位在两种RNA二级结构中的双螺旋区域。这些结果证实了核糖体RNA对核糖体功能的重要性,并有助于确定构成大肠杆菌核糖体亚基界面的RNA结构域。