Spitnik-Elson P, Elson D, Avital S, Abramowitz R
Nucleic Acids Res. 1985 Jul 11;13(13):4719-38. doi: 10.1093/nar/13.13.4719.
We have attempted to identify long-range interactions in the tertiary structure of RNA in the E. coli 30 S ribosome. Native subunits were cleaved with ribonuclease and separated into nucleoprotein fragments which were deproteinized and fractionated into multi-oligonucleotide complexes under conditions intended to preserve RNA-RNA interactions. The final products were denatured by urea and heat and their constituent oligonucleotides resolved and sequenced. Many complexes contained complementary sequences known to be bound together in the RNA secondary structure, attesting to the validity of the technique. Other co-migrating oligonucleotides, not joined in the secondary structure, contained mutually complementary sequences in locations that allow base-pairing interaction without disrupting pre-existing secondary structure. In seven instances the complementary relationship was found to have been preserved during phylogenetic diversification.
我们试图确定大肠杆菌30S核糖体中RNA三级结构的长程相互作用。天然亚基用核糖核酸酶切割,分离成核蛋白片段,这些片段经过脱蛋白处理,并在旨在保留RNA-RNA相互作用的条件下分级分离成多寡核苷酸复合物。最终产物用尿素和加热使其变性,其组成的寡核苷酸被分离并测序。许多复合物包含已知在RNA二级结构中结合在一起的互补序列,证明了该技术的有效性。其他共同迁移的寡核苷酸,在二级结构中未连接,在允许碱基配对相互作用而不破坏预先存在的二级结构的位置包含相互互补的序列。在七个实例中,发现互补关系在系统发育多样化过程中得以保留。