一项对大肠杆菌16S核糖体RNA 3'端的碳-13核磁共振研究，该3'端在m6(2)Am6(2)A序列的甲基中被碳-13特异性标记。
A carbon-13 nuclear magnetic resonance study of the 3'-terminus of 16S ribosomal RNA of Escherichia coli specifically labeled with carbon-13 in the methylgroups of the m6(2)Am6(2)A sequence.
作者信息
Van Charldorp R, Verhoeven J J, Van Knippenberg P H, Haasnoot C A, Hilbers C W
出版信息
Nucleic Acids Res. 1982 Jul 24;10(14):4237-45. doi: 10.1093/nar/10.14.4237.
Abstract
30S ribosomes were isolated from a kasugamycin resistant mutant of E. coli that lacks methylgroups on two adjacent adenines in 16S ribosomal RNA. These ribosomes were methylated in vitro with a purified methylating enzyme and 5-S-adenosyl-(13C-methyl)-L-methionine chloride ((13C-methyl)-SAM) as methyldonor. After in situ cleavage of the 16S ribosomal RNA by the bacteriocin cloacin DF13, the 49 nucleotide fragment from the 3'-end of the RNA was isolated. The carbon-13 nuclear magnetic resonance spectra of the fragment at various temperatures were compared with those of 6-N-dimethyladenosine (m6(2)A) and 6-N-dimethyladenylyl-(3' leads to 5')-6-N-dimethyladenosine (m6(2)Am6(2)A). The data show that the two methylated adenines, which are part of a four membered hairpin loop, show a strong tendency to be stacked in analogy to the dinucleotide m6(2)Am6(2).
摘要
从一株对春雷霉素耐药的大肠杆菌突变体中分离出30S核糖体，该突变体的16S核糖体RNA中两个相邻腺嘌呤上缺乏甲基基团。这些核糖体在体外使用纯化的甲基化酶和5'-S-腺苷基-(13C-甲基)-L-甲硫氨酸氯化物((13C-甲基)-SAM)作为甲基供体进行甲基化。在用细菌素cloacin DF13对16S核糖体RNA进行原位切割后，从RNA的3'端分离出49个核苷酸的片段。将该片段在不同温度下的碳-13核磁共振谱与6-N-二甲基腺苷(m6(2)A)和6-N-二甲基腺苷基-(3'→5')-6-N-二甲基腺苷(m6(2)Am6(2)A)的谱进行比较。数据表明，作为四元发夹环一部分的两个甲基化腺嘌呤，与二核苷酸m6(2)Am6(2)类似，表现出强烈的堆积倾向。
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