Trempe M R, Ohgi K, Glitz D G
J Biol Chem. 1982 Aug 25;257(16):9822-9.
The minor nucleoside 7-methylguanosine occurs in Escherichia coli 16 S ribosomal RNA at a single site. High pressure liquid chromatographic analysis shows that a single residue of 7-methylguanosine is also present in chloroplast 16 S ribosomal RNA, presumably at an analogous position in the sequence. Antibodies to 7-methylguanosine were induced in rabbits and shown to be highly specific for the intact methylated base. These antibodies were reacted with 30 S ribosomal subunits from E. coli and from the chloroplasts of Alaskan peas. These two types of ribosome have been shown to be topographically similar (Trempe, M. R., and Glitz, D. G. (1981) J. Biol. Chem. 256, 11873-11879). Electron microscopy of the subunit-antibody complexes showed similar subunit-IgG monomers and antibody-linked subunit dimers. In greater than 95% of the complexes observed for each type of ribosome, antibody contact was consistent with a single binding site, which places 7-methylguanosine near the junction of the upper one-third and lower two-thirds of the subunit and maximally distant from the platform. The analogous localization in both E. coli and chloroplast 30 S ribosomal subunits lends support to their proposed common evolutionary origin.
次要核苷7-甲基鸟苷在大肠杆菌16S核糖体RNA中仅存在于一个位点。高压液相色谱分析表明,叶绿体16S核糖体RNA中也存在一个7-甲基鸟苷残基,推测在序列中的类似位置。在兔子体内诱导产生了针对7-甲基鸟苷的抗体,并证明其对完整的甲基化碱基具有高度特异性。这些抗体与来自大肠杆菌和阿拉斯加豌豆叶绿体的30S核糖体亚基反应。这两种类型的核糖体在拓扑结构上已被证明是相似的(特伦普,M.R.,和格利茨,D.G.(1981年)《生物化学杂志》256,11873 - 11879)。亚基 - 抗体复合物的电子显微镜观察显示出相似的亚基 - IgG单体和抗体连接的亚基二聚体。在每种类型核糖体观察到的超过95%的复合物中,抗体接触与单个结合位点一致,该位点将7-甲基鸟苷置于亚基上三分之一和下三分之二交界处附近,且距离平台最远。大肠杆菌和叶绿体30S核糖体亚基中的类似定位支持了它们共同的进化起源假说。