Douthwaite S, Garrett R A
Biochemistry. 1981 Dec 8;20(25):7301-7. doi: 10.1021/bi00528a039.
The structures of 5S ribosomal RNAs from Escherichia coli and Bacillus stearothermophilus were examined by using ribonucleases A, T1, and T2 and a double helix specific cobra venom ribonuclease. By using both 5' and 3'-32P-end labeling methods and selecting for digested but intact 5S RNA molecules, we were able to distinguish between primary and secondary cutting positions and also to establish the relative degree of cutting. The data reveal the predicted similarities of the higher order structure in the two RNAs but also demonstrate a few significant differences. The data also provide direct evidence for three of the helical regions of the Fox and Woese model of 5S RNA [Fox, G. E., & Woese, C. (1975) Nature (London) 256, 505] and support other important structural features which include a nucleotide looped out from a helical region which has been proposed as a recognition site for protein L18.
利用核糖核酸酶A、T1、T2以及一种双链螺旋特异性眼镜蛇毒核糖核酸酶,对来自大肠杆菌和嗜热栖热芽孢杆菌的5S核糖体RNA的结构进行了研究。通过使用5'和3'-32P末端标记方法,并选择经过消化但完整的5S RNA分子,我们能够区分一级和二级切割位点,并确定相对切割程度。数据揭示了两种RNA在高级结构上的预测相似性,但也显示出一些显著差异。这些数据还为Fox和Woese的5S RNA模型[Fox, G. E., & Woese, C.(1975) Nature (London) 256, 505]的三个螺旋区域提供了直接证据,并支持其他重要的结构特征,其中包括一个从螺旋区域中突出的核苷酸,该区域被认为是蛋白质L18的识别位点。