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在 中鉴定核基因的 U 到 C RNA 编辑事件的全基因组分析

Genome-Wide Identification of U-To-C RNA Editing Events for Nuclear Genes in .

机构信息

Area of Bioscience and Biotechnology, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi City, Ishikawa 923-1292, Japan.

Division of Transdisciplinary Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi City, Ishikawa 923-1292, Japan.

出版信息

Cells. 2021 Mar 12;10(3):635. doi: 10.3390/cells10030635.

Abstract

Cytosine-to-Uridine (C-to-U) RNA editing involves the deamination phenomenon, which is observed in animal nucleus and plant organelles; however, it has been considered the U-to-C is confined to the organelles of limited non-angiosperm plant species. Although previous RNA-seq-based analysis implied U-to-C RNA editing events in plant nuclear genes, it has not been broadly accepted due to inadequate confirmatory analyses. Here we examined the U-to-C RNA editing in Arabidopsis tissues at different developmental stages of growth. In this study, the high-throughput RNA sequencing (RNA-seq) of 12-day-old and 20-day-old Arabidopsis seedlings was performed, which enabled transcriptome-wide identification of RNA editing sites to analyze differentially expressed genes (DEGs) and nucleotide base conversions. The results showed that DEGs were expressed to higher levels in 12-day-old seedlings than in 20-day-old seedlings. Additionally, pentatricopeptide repeat (PPR) genes were also expressed at higher levels, as indicated by the logFC values. RNA-seq analysis of 12-day- and 20-day-old Arabidopsis seedlings revealed candidates of U-to-C RNA editing events. Sanger sequencing of both DNA and cDNA for all candidate nucleotide conversions confirmed the seven U-to-C RNA editing sites. This work clearly demonstrated presence of U-to-C RNA editing for nuclear genes in Arabidopsis, which provides the basis to study the mechanism as well as the functions of the unique post-transcriptional modification.

摘要

胞嘧啶向尿嘧啶(C-to-U)RNA 编辑涉及脱氨酶现象,这种现象在动物细胞核和植物细胞器中都有观察到;然而,人们一直认为 U-to-C 仅限于有限的非被子植物物种的细胞器。尽管之前基于 RNA-seq 的分析暗示了植物核基因中的 U-to-C RNA 编辑事件,但由于缺乏确认性分析,这一观点并未得到广泛接受。在这里,我们研究了不同生长发育阶段的拟南芥组织中的 U-to-C RNA 编辑。在这项研究中,对 12 天和 20 天龄的拟南芥幼苗进行了高通量 RNA 测序(RNA-seq),这使得能够在全转录组范围内识别 RNA 编辑位点,以分析差异表达基因(DEGs)和核苷酸碱基转换。结果表明,在 12 天龄的幼苗中,DEGs 的表达水平高于 20 天龄的幼苗。此外,五肽重复(PPR)基因的表达水平也更高,这一点从 logFC 值可以看出。对 12 天和 20 天龄拟南芥幼苗的 RNA-seq 分析揭示了 U-to-C RNA 编辑事件的候选者。对所有候选核苷酸转换的 DNA 和 cDNA 的 Sanger 测序均证实了七个 U-to-C RNA 编辑位点的存在。这项工作清楚地表明了拟南芥核基因中存在 U-to-C RNA 编辑,为研究这种独特的转录后修饰的机制和功能提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c258/8001311/7f21cd1ba637/cells-10-00635-g001.jpg

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