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通过碱基脱氨作用控制基因表达:小麦线粒体中的RNA编辑实例

Control of gene expression by base deamination: the case of RNA editing in wheat mitochondria.

作者信息

Blanc V, Jordana X, Litvak S, Araya A

机构信息

Institut de Biochimie et Génétique Cellulaires, IBGC-CNRS, Bordeaux, France.

出版信息

Biochimie. 1996;78(6):511-7. doi: 10.1016/0300-9084(96)84757-2.

Abstract

The term 'RNA editing' was used for the first time in 1986 to describe the process of uridylate insertion into trypanosomal mitochondrial transcripts. Since then, the term has been used more generally to describe a large variety of processes involving base insertions, deletions and conversions that generate RNAs with a primary sequence different to those encoded by the gene. RNA editing has been observed in the mitochondrial fraction of trypanosomes, plants and other organisms, in the animal nuclear fraction in the case of the apolipoprotein B and glutamate brain receptors mRNAs as well as in viruses like paramyxovirus, hepatitis delta and probably HIV. The role of cytidine and adenine deamination leading to C to U and A to I transitions has became pivotal to explain this process by base conversion. In this review we will focus mainly on the work performed in our group on plant mitochondria and more specifically on the mechanism and the functional significance of RNA editing in wheat organelles. The original contributions of our laboratory in this field are: i) showing that RNA editing is reflected at the protein level; ii) settling three in vitro systems to assay C to U conversion using a wheat mitochondrial lysate as source of enzymes and factors, and unedited mRNA from the same source, as substrate; iii) determination by double labelling of the unedited substrate that RNA editing in wheat mitochondria occurs via a deamination step; and iv) that introducing unedited proteins in the mitochondria of transgenic plants leads to the emergence of cytoplasmic male sterility supporting the idea that the role of this process is to produce functional proteins. Using the antisense approach in transgenic plants we were able to obtain a significant male fertility restoration.

摘要

“RNA编辑”一词于1986年首次被用于描述尿苷酸插入锥虫线粒体转录本的过程。从那时起,该术语被更广泛地用于描述各种各样的过程,这些过程涉及碱基插入、缺失和转换,从而产生与基因编码的初级序列不同的RNA。在锥虫、植物和其他生物体的线粒体部分,在载脂蛋白B和谷氨酸脑受体mRNA的动物细胞核部分以及在副粘病毒、丁型肝炎病毒和可能的HIV等病毒中都观察到了RNA编辑。胞嘧啶和腺嘌呤脱氨基导致C到U和A到I转换的作用已成为通过碱基转换解释这一过程的关键。在这篇综述中,我们将主要关注我们小组在植物线粒体方面所做的工作,更具体地说是小麦细胞器中RNA编辑的机制和功能意义。我们实验室在这一领域的原创性贡献包括:i)表明RNA编辑在蛋白质水平上得到体现;ii)建立三个体外系统,以小麦线粒体裂解物作为酶和因子的来源,以及来自同一来源的未编辑mRNA作为底物,来检测C到U的转换;iii)通过对未编辑底物的双重标记确定小麦线粒体中的RNA编辑是通过脱氨基步骤发生的;iv)在转基因植物的线粒体中引入未编辑的蛋白质会导致细胞质雄性不育的出现,这支持了这一过程的作用是产生功能性蛋白质的观点。利用转基因植物中的反义方法,我们能够显著恢复雄性育性。

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