IZMB - Institut für Zelluläre und Molekulare Botanik, Abt. Molekulare Evolution, University of Bonn, Kirschallee 1, 53115, Bonn, Germany.
Commun Biol. 2019 Mar 1;2:85. doi: 10.1038/s42003-019-0328-3. eCollection 2019.
RNA editing converting cytidines into uridines is a hallmark of gene expression in land plant chloroplasts and mitochondria. Pentatricopeptide repeat (PPR) proteins have a key role in target recognition, but the functional editosome in the plant organelles has remained elusive. Here we show that individual DYW-type PPR proteins alone can perform efficient C-to-U editing in reproducing the moss mitochondrial editing. Single amino acid exchanges in the DYW domain abolish RNA editing, confirming it as the functional cytidine deaminase. The modification of RNA targets and the identification of numerous off-targets in the transcriptome reveal nucleotide identities critical for RNA recognition and cytidine conversion. The straightforward amenability of the new setup will accelerate future studies on RNA target recognition through PPRs, on the C-to-U editing deamination machinery and towards future establishment of transcript editing in other genetic systems.
RNA 编辑将胞嘧啶转化为尿嘧啶是陆地植物叶绿体和线粒体基因表达的一个标志。五肽重复（PPR）蛋白在靶标识别中起着关键作用，但植物细胞器中的功能性编辑体仍然难以捉摸。在这里，我们表明单个 DYW 型 PPR 蛋白本身就可以在复制苔藓线粒体编辑中进行有效的 C 到 U 编辑。DYW 结构域中的单个氨基酸交换会使 RNA 编辑失效，从而证实它是具有功能的胞嘧啶脱氨酶。RNA 靶标的修饰和在转录组中鉴定出大量的脱靶表明，核苷酸的同一性对于 RNA 识别和胞嘧啶转化至关重要。新方法的简单易用性将加速未来通过 PPR 对 RNA 靶标识别、C 到 U 编辑脱氨酶机制以及在其他遗传系统中建立转录编辑的研究。
