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全基因组鉴定和.UTR-内含子的特征分析

Genome-Wide Identification and Characterization of UTR-Introns of .

机构信息

College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China.

Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China.

出版信息

Int J Mol Sci. 2020 Apr 27;21(9):3088. doi: 10.3390/ijms21093088.

Abstract

Introns exist not only in coding sequences (CDSs) but also in untranslated regions (UTRs) of a gene. Recent studies in animals and model plants such as have revealed that the UTR-introns (UIs) are widely presented in most genomes and involved in regulation of gene expression or RNA stability. In the present study, we identified introns at both 5'UTRs (5UIs) and 3'UTRs (3UIs) of sweet orange genes, investigated their size and nucleotide distribution characteristics, and explored the distribution of -elements in the UI sequences. Functional category of genes with predicted UIs were further analyzed using GO, KEGG, and PageMan enrichment. In addition, the organ-dependent splicing and abundance of selected UI-containing genes in root, leaf, and stem were experimentally determined. Totally, we identified 825 UI- and 570 3UI-containing transcripts, corresponding to 617 and 469 genes, respectively. Among them, 74 genes contain both 5UI and 3UI. Nucleotide distribution analysis showed that 5UI distribution is biased at both ends of 5'UTR whiles 3UI distribution is biased close to the start site of 3'UTR. elements analysis revealed that 5UI and 3UI sequences were rich of promoter-enhancing related elements, indicating that they might function in regulating the expression through them. Function enrichment analysis revealed that genes containing 5UI are significantly enriched in the RNA transport pathway. While, genes containing 3UI are significantly enriched in splicesome. Notably, many and the were identified to be 3UI-containing. RT-PCR result confirmed the existence of UIs in the eight selected gene transcripts whereas alternative splicing events were found in some of them. Meanwhile, qRT-PCR result showed that UIs were differentially expressed among organs, and significant correlation was found between some genes and their UIs, for example: The expression of and its 3UI was significantly negative correlated. This is the first report about the UIs in sweet orange from genome-wide level, which could provide evidence for further understanding of the role of UIs in gene expression regulation.

摘要

内含子不仅存在于编码序列 (CDS) 中,也存在于基因的非翻译区 (UTR) 中。最近在动物和模式植物中的研究表明,UTR 内含子 (UIs) 广泛存在于大多数基因组中,并参与基因表达或 RNA 稳定性的调控。在本研究中,我们鉴定了甜橙基因的 5'UTR(5UIs)和 3'UTR(3UIs)中的内含子,研究了它们的大小和核苷酸分布特征,并探讨了 UI 序列中 - 元件的分布。使用 GO、KEGG 和 PageMan 富集进一步分析了具有预测 UI 的基因的功能类别。此外,还通过实验确定了在根、叶和茎中选择的含有 UI 的基因的器官依赖性剪接和丰度。总共鉴定了 825 个 UI 和 570 个 3UI 转录本,分别对应于 617 个和 469 个基因。其中,74 个基因同时含有 5UI 和 3UI。核苷酸分布分析表明,5UI 分布偏向 5'UTR 的两端,而 3UI 分布偏向 3'UTR 的起始位点。- 元件分析表明,5UI 和 3UI 序列富含启动子增强相关元件,表明它们可能通过这些元件发挥作用调节表达。功能富集分析表明,含有 5UI 的基因在 RNA 运输途径中显著富集。而含有 3UI 的基因在剪接体中显著富集。值得注意的是,许多 和 被鉴定为含有 3UI。RT-PCR 结果证实了在所选择的 8 个基因转录本中存在 UI,而在其中一些转录本中发现了可变剪接事件。同时,qRT-PCR 结果表明,UI 在器官间存在差异表达,并且一些基因与其 UI 之间存在显著相关性,例如: 及其 3UI 的表达呈显著负相关。这是从全基因组水平上首次报道甜橙中的 UI,为进一步了解 UI 在基因表达调控中的作用提供了证据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac5f/7247714/86d4db0e0db3/ijms-21-03088-g001.jpg

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