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节节麦叶绿体转录本中RNA编辑位点的鉴定与分析

Identification and Analysis of RNA Editing Sites in the Chloroplast Transcripts of Aegilops tauschii L.

作者信息

Wang Mengxing, Liu Hui, Ge Lingqiao, Xing Guangwei, Wang Meng, Weining Song, Nie Xiaojun

机构信息

State Key Laboratory of Crop Stress Biology in Arid Areas, College of Agronomy, Northwest Agriculture and Forestry University, Yangling 712100, Xianyang, China.

出版信息

Genes (Basel). 2016 Dec 30;8(1):13. doi: 10.3390/genes8010013.

Abstract

RNA editing is an important way to convert cytidine (C) to uridine (U) at specific sites within RNA molecules at a post-transcriptional level in the chloroplasts of higher plants. Although it has been systematically studied in many plants, little is known about RNA editing in the wheat D genome donor L. Here, we investigated the chloroplast RNA editing of and compared it with other wheat relatives to trace the evolution of wheat. Through bioinformatics prediction, a total of 34 C-to-U editing sites were identified, 17 of which were validated using RT-PCR product sequencing. Furthermore, 60 sites were found by the RNA-Seq read mapping approach, 24 of which agreed with the prediction and six were validated experimentally. The editing sites were biased toward tCn or nCa trinucleotides and 5'-pyrimidines, which were consistent with the flanking bases of editing sites of other seed plants. Furthermore, the editing events could result in the alteration of the secondary structures and topologies of the corresponding proteins, suggesting that RNA editing might impact the function of target genes. Finally, comparative analysis found some evolutionarily conserved editing sites in wheat and two species-specific sites were also obtained. This study is the first to report on RNA editing in L, which not only sheds light on the evolution of wheat from the point of view of RNA editing, but also lays a foundation for further studies to identify the mechanisms of C-to-U alterations.

摘要

RNA编辑是高等植物叶绿体中在转录后水平将RNA分子特定位点的胞嘧啶(C)转化为尿嘧啶(U)的重要方式。尽管在许多植物中已对其进行了系统研究,但对于小麦D基因组供体节节麦中的RNA编辑却知之甚少。在此,我们研究了节节麦的叶绿体RNA编辑,并将其与其他小麦近缘种进行比较以追溯小麦的进化。通过生物信息学预测,共鉴定出34个C到U的编辑位点,其中17个通过RT-PCR产物测序得到验证。此外,通过RNA-Seq读段映射方法发现了60个位点,其中24个与预测结果一致,6个通过实验得到验证。编辑位点偏向于tCn或nCa三核苷酸以及5'-嘧啶,这与其他种子植物编辑位点的侧翼碱基一致。此外,编辑事件可能导致相应蛋白质二级结构和拓扑结构的改变,表明RNA编辑可能影响靶基因的功能。最后,比较分析发现了小麦中一些进化上保守的编辑位点,还获得了两个物种特有的位点。本研究首次报道了节节麦中的RNA编辑,这不仅从RNA编辑的角度揭示了小麦的进化,也为进一步研究鉴定C到U改变的机制奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f0fd/5295008/f89d9ce4a63d/genes-08-00013-g001.jpg

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