Plant-type pentatricopeptide repeat proteins with a DYW domain drive C-to-U RNA editing in Escherichia coli
Plant-type pentatricopeptide repeat proteins with a DYW domain drive C-to-U RNA editing in Escherichia coli
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DOI：
10.1038/s42003-019-0328-3
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发表时间：
2019-03-01
影响因子：
5.9
通讯作者：
Schallenberg-Ruedinger, Mareike
中科院分区：
文献类型：
--
作者：
Oldenkott, Bastian;Yang, Yingying;Schallenberg-Ruedinger, Mareike
将胞苷转换为尿苷的RNA编辑是陆地植物叶绿体和线粒体中基因表达的标志。五肽重复序列（PPR）蛋白在靶标识别中起着关键作用，但植物细胞器中功能性编辑体的研究一直是个谜。在这里，我们表明，单独的小立碗藓DYW型PPR蛋白可以在大肠杆菌中进行有效的C-to-U编辑，重现苔藓线粒体编辑。DYW结构域中的单个氨基酸交换消除了RNA编辑，证实其为功能性胞苷脱氨酶。对RNA靶点的修饰和在大肠杆菌中大量脱靶的鉴定是目前研究的热点。大肠杆菌转录组揭示了对RNA识别和胞苷转化至关重要的核苷酸同一性。新E.大肠杆菌的建立将加速未来通过PPR识别RNA靶点的研究，C-to-U编辑脱氨基机制，以及未来在其他遗传系统中建立转录本编辑。
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 Physcomitrella patens DYW-type PPR proteins alone can perform efficient C-to-U editing in Escherichia coli 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 E. coli transcriptome reveal nucleotide identities critical for RNA recognition and cytidine conversion. The straightforward amenability of the new E. coli 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.