Blanc V, Litvak S, Araya A
Institut de Biochimie et Génétique Cellulaires, IBGC-CNRS, Bordeaux, France.
FEBS Lett. 1995 Oct 2;373(1):56-60. doi: 10.1016/0014-5793(95)00991-h.
Most if not all mitochondrial messenger RNAs from seed plants undergo a post-transcriptional modification (RNA editing) involving the conversion of some cytidine residues to uridine. Using a molecular hybridization approach, an in vitro RNA editing system, able to faithfully reproduce the in vivo observed C to U changes of subunit 9 (atp9) of wheat mitochondrial ATP synthase mRNA, has been described [Araya et al. (1992) Proc. Natl. Acad. Sci. USA 89, 1040-1044]. In this work we extend these studies to better understand the biochemical mechanism of this process. RNA editing was analysed by P1 nuclease digestion of the reaction product followed by thin layer chromatography. Experiments performed with unedited [3H]RNA labelled on the base and with unedited [32P]RNA labelled at the alpha-phosphate of cytidine residues, indicate that plant mitochondrial RNA editing operates through a deamination mechanism.
大多数(如果不是全部的话)种子植物的线粒体信使核糖核酸会经历一种转录后修饰(RNA编辑),该修饰涉及将一些胞嘧啶残基转变为尿嘧啶。利用分子杂交方法,已经描述了一种体外RNA编辑系统,它能够如实地重现体内观察到的小麦线粒体ATP合酶信使核糖核酸亚基9(atp9)的胞嘧啶(C)到尿嘧啶(U)的变化 [阿雷亚等人(1992年)《美国国家科学院院刊》89, 1040 - 1044]。在这项工作中,我们扩展这些研究以更好地理解这一过程的生化机制。通过对反应产物进行P1核酸酶消化,然后进行薄层色谱分析来研究RNA编辑。用在碱基上标记的未编辑的[³H]RNA以及在胞嘧啶残基的α - 磷酸上标记的未编辑的[³²P]RNA所进行的实验表明,植物线粒体RNA编辑是通过脱氨机制进行的。