Wiener L, Brimacombe R
Nucleic Acids Res. 1987 May 11;15(9):3653-70. doi: 10.1093/nar/15.9.3653.
14C-labelled proteins from E. coli 30S ribosomal subunits were isolated by HPLC, and selected groups of these proteins were reconstituted with 32P-labelled 16S RNA. The isolated reconstituted particles were partially digested with ribonuclease A, and the RNA fragments protected by the proteins were separated by gel electrophoresis and subjected to sequence analysis. Protein S7 alone gave no protected fragments, but S7 together with S14 and S19 protected an RNA region comprising the sequences 936-965, 972-1030, 1208-1262 and 1285-1379 of the 16S RNA. Addition of increasing amounts of protein S9 to the S7/S14/S19 particle resulted in a parallel increase in the protection of the hairpin loop between bases 1262 and 1285. The results are discussed in terms of the three-dimensional folding of 16S RNA in the 30S subunit.
通过高效液相色谱法从大肠杆菌30S核糖体亚基中分离出14C标记的蛋白质,并将这些蛋白质的选定组与32P标记的16S RNA进行重组。分离出的重组颗粒用核糖核酸酶A进行部分消化,受蛋白质保护的RNA片段通过凝胶电泳分离并进行序列分析。单独的蛋白质S7没有产生受保护的片段,但S7与S14和S19一起保护了一个RNA区域,该区域包含16S RNA的936 - 965、972 - 1030、1208 - 1262和1285 - 1379序列。向S7/S14/S19颗粒中添加越来越多的蛋白质S9导致1262和1285碱基之间发夹环的保护作用平行增加。根据30S亚基中16S RNA的三维折叠对结果进行了讨论。