Stiege W, Stade K, Schüler D, Brimacombe R
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin-Dahlem, FRG.
Nucleic Acids Res. 1988 Mar 25;16(6):2369-88. doi: 10.1093/nar/16.6.2369.
Poly(A) can be cross-linked to E. coli 70S ribosomes in the presence of tRNALys by mild ultraviolet irradiation. The cross-linking reaction is exclusively with the 30S subunit, and involves primarily the RNA moiety. Following a partial nuclease digestion, cross-linked complexes containing poly(A) and fragments of the 16S RNA were isolated by affinity chromatography on oligo(dT)-cellulose. The complexes were purified by gel electrophoresis and subjected to oligonucleotide analysis, which revealed a single cross-link site within positions 1394-1399 of the 16S RNA. The same pattern of cross-linking, at about one-fifth of the intensity, was observed in the absence of tRNALys. The cross-link site to poly(A), together with other sites in the 16S RNA that have been implicated in ribosomal function, is discussed in the framework of our recent model for the three-dimensional structure of 16S RNA; all of the functional sites are clustered together in two distinct groups in the model.
在赖氨酸转运RNA（tRNALys）存在的情况下，通过温和的紫外线照射，聚腺苷酸（Poly(A)）可与大肠杆菌70S核糖体发生交联。这种交联反应仅发生在30S亚基上，并且主要涉及RNA部分。经过部分核酸酶消化后，通过在寡聚（dT）-纤维素上进行亲和层析，分离出含有聚腺苷酸（Poly(A)）和16S RNA片段的交联复合物。通过凝胶电泳对复合物进行纯化，并进行寡核苷酸分析，结果显示在16S RNA的1394 - 1399位之间存在一个单一的交联位点。在没有赖氨酸转运RNA（tRNALys）的情况下，观察到相同的交联模式，但其强度约为有tRNALys时的五分之一。在我们最近关于16S RNA三维结构的模型框架内，讨论了聚腺苷酸（Poly(A)）的交联位点以及16S RNA中其他与核糖体功能相关的位点；在该模型中，所有功能位点聚集在两个不同的组中。
