State Key Laboratory of Hybrid Rice; Engineering Research Center for Plant Biotechnology and Germplasm Utilization of Ministry of Education, College of Life Sciences, Wuhan University, Wuhan, China.
J Exp Bot. 2018 May 25;69(12):2923-2936. doi: 10.1093/jxb/ery108.
In flowering plants, various RNA editing events occur in the mitochondria and chloroplasts as part of post-transcriptional processes. Although several pentatricopeptide repeat (PPR) proteins and multiple organellar RNA editing factors (MORFs) have been identified as RNA editing factors, the underlying mechanism of PPRs and the cooperation among these proteins are still obscure. Here, we identified a rice dual-localized PPR protein, OsPGL1. The loss of function of OsPGL1 resulted in defects in both chloroplast RNA editing of ndhD-878 and mitochondrial RNA editing of ccmFc-543, both of which could be restored in transgenic complementation lines. Despite synonymous editing of ccmFc-543, the loss of editing of ndhD-878 caused a failed conversion of serine to leucine, leading to chloroplast dysfunction and defects in the photosynthetic complex; the results of additional experiments demonstrated that OsPGL1 directly binds to both transcripts. Interactions between three OsMORFs (OsMORF2/8/9) and OsPGL1 both in vitro and in vivo were confirmed, implying that OsPGL1 functions in RNA editing via an editosome. These findings also suggested that OsMORFs assist with and contribute to a flexible PPR-RNA recognition model during RNA editing. These results indicate that, in cooperation with PPRs, OsPGL1 is required for RNA editing. In addition, our study provides new insights into the relationship between RNA editing and plant development.
在开花植物中,各种 RNA 编辑事件发生在线粒体和叶绿体中,作为转录后过程的一部分。尽管已经鉴定出几种五肽重复(PPR)蛋白和多个细胞器 RNA 编辑因子(MORFs)作为 RNA 编辑因子,但 PPR 蛋白的潜在机制以及这些蛋白之间的合作仍然不清楚。在这里,我们鉴定了一个水稻双重定位的 PPR 蛋白,OsPGL1。OsPGL1 功能丧失导致叶绿体 RNA 编辑 ndhD-878 和线粒体 RNA 编辑 ccmFc-543 的缺陷,这两种缺陷都可以在转基因互补系中得到恢复。尽管 ccmFc-543 的编辑是同义的,但 ndhD-878 编辑的缺失导致丝氨酸到亮氨酸的转换失败,导致叶绿体功能障碍和光合复合物缺陷;额外的实验结果表明,OsPGL1 直接结合这两个转录本。体外和体内都证实了三个 OsMORFs(OsMORF2/8/9)和 OsPGL1 之间的相互作用,暗示 OsPGL1 通过编辑体发挥 RNA 编辑功能。这些发现还表明,OsMORFs 在 RNA 编辑过程中协助并促成了一种灵活的 PPR-RNA 识别模型。这些结果表明,在与 PPR 蛋白合作的情况下,OsPGL1 是 RNA 编辑所必需的。此外,我们的研究为 RNA 编辑与植物发育之间的关系提供了新的见解。