Wrede P, Wurst R, Vournakis J, Rich A
J Biol Chem. 1979 Oct 10;254(19):9608-16.
The susceptibility of yeast tRNAPhe and Escherichia coli tRNA2Glu to digestion by nucleases Tl and Sl are examined in a variety of environments, and the results are interpreted in view of the available three-dimensional structural information. Significant differences are found in the digestion pattern of the two tRNAs using the guanosine-specific Tl nuclease. In particular, differences are seen due to varying the type of salts in the environment. However, the Sl nuclease results on the two tRNAs do not differ greatly. E. coli tRNA2Glu is known to exist in two different conformations. Nuclease digestion results are presented revealing differences which make it possible to draw some inferences about the structural differences in these two conformations. In carrying out these analyses, the tRNA molecules are labeled either by putting 32P at the 5'-end of the molecular or by adding 32P-labeled pCp at the 3'-end. It is found that both yeast tRNAPhe and E. coli tRNA2Glu have modified Tl nuclease digestion patterns when pCp is added at the 3'-end of the molecule.
在多种环境下检测了酵母苯丙氨酸转运核糖核酸(tRNAPhe)和大肠杆菌谷氨酰胺转运核糖核酸2(tRNA2Glu)对核酸酶Tl和Sl消化的敏感性,并根据现有的三维结构信息对结果进行了解释。使用鸟苷特异性核酸酶Tl时,发现两种转运核糖核酸的消化模式存在显著差异。特别是,由于环境中盐的类型不同而出现了差异。然而,两种转运核糖核酸的核酸酶Sl消化结果差异不大。已知大肠杆菌tRNA2Glu存在两种不同的构象。给出的核酸酶消化结果揭示了差异,从而有可能对这两种构象的结构差异做出一些推断。在进行这些分析时,通过在分子的5'端放置32P或在3'端添加32P标记的对氯苯磷酸(pCp)来标记转运核糖核酸分子。发现当在分子的3'端添加pCp时,酵母tRNAPhe和大肠杆菌tRNA2Glu都有改变的核酸酶Tl消化模式。