Shanghai Key Laboratory of Bio-Energy Crops, School of Life Sciences, Shanghai University, Shanghai 200444, China.
Shanghai Key Laboratory of Bio-Energy Crops, School of Life Sciences, Shanghai University, Shanghai 200444, China; Coordinated Crop Biology Research Center (CBRC), Beijing 100193, China.
Mol Plant. 2017 Mar 6;10(3):427-441. doi: 10.1016/j.molp.2016.08.008. Epub 2016 Sep 3.
In higher plants, the splicing of organelle-encoded mRNA involves a complex collaboration with nuclear-encoded proteins. Pentatricopeptide repeat (PPR) proteins have been implicated in these RNA-protein interactions. In this study, we performed the cloning and functional characterization of maize Defective kernel 35 (Dek35). The dek35-ref mutant is a lethal-seed mutant with developmental deficiency. Dek35 was cloned through Mutator tag isolation and further confirmed by four additional independent mutant alleles. Dek35 encodes an P-type PPR protein that targets the mitochondria. The dek35 mutation causes significant reduction in the accumulation of DEK35 proteins and reduced splicing efficiency of mitochondrial nad4 intron 1. Analysis of mitochondrial complex in dek35 immature seeds indicated severe deficiency in the complex I assembly and NADH dehydrogenase activity. Transcriptome analysis of dek35 endosperm revealed enhanced expression of genes involved in the alternative respiratory pathway and extensive differentially expressed genes related to mitochondrial function and activity. Collectively, these results indicate that Dek35 encodes an PPR protein that affects the cis-splicing of mitochondrial nad4 intron 1 and is required for mitochondrial function and seed development.
在高等植物中,细胞器编码的 mRNA 的剪接涉及与核编码蛋白的复杂协作。五肽重复(PPR)蛋白已被牵涉到这些 RNA-蛋白相互作用中。在这项研究中,我们进行了玉米缺陷核 35(Dek35)的克隆和功能表征。dek35-ref 突变体是一种致死性种子突变体,表现出发育缺陷。通过 Mutator 标签分离克隆 dek35,并通过另外四个独立的突变等位基因进一步证实。Dek35 编码一种靶向线粒体的 P 型 PPR 蛋白。dek35 突变导致 DEK35 蛋白积累显著减少,线粒体 nad4 内含子 1 的剪接效率降低。对 dek35 不成熟种子中线粒体复合物的分析表明,复合物 I 的组装和 NADH 脱氢酶活性严重缺乏。对 dek35 胚乳的转录组分析表明,参与替代呼吸途径的基因表达增强,与线粒体功能和活性相关的大量差异表达基因。总之,这些结果表明 Dek35 编码一种 PPR 蛋白,它影响线粒体 nad4 内含子 1 的顺式剪接,并且是线粒体功能和种子发育所必需的。