Differential Binding of Three Major Human ADAR Isoforms to Coding and Long Non-Coding Transcripts.
Differential Binding of Three Major Human ADAR Isoforms to Coding and Long Non-Coding Transcripts.
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DOI：
10.3390/genes8020068
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发表时间：
2017-02-11
期刊：
影响因子：
3.5
通讯作者：
Ui-Tei K
中科院分区：
文献类型：
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作者：
Galipon J;Ishii R;Suzuki Y;Tomita M;Ui-Tei K
通过腺苷脱胺为肌苷的RNA编辑是一个进化上保守的过程，涉及许多细胞途径，从选择性剪接到miRNA靶向。在人类中，至少有三种主要的腺苷脱氨酶作用于RNA (ADARs): ADAR1-p150， ADAR1-p110和ADAR2。然而，前两个基因源于选择性剪接，因此目前不可能在不敲除ADAR1-p150表达的情况下删除ADAR1-p110。此外，ADARs的表达水平在不同的细胞类型中差异很大，目前还没有研究系统地探讨这些亚型对细胞转录组的影响。在本研究中，对绿色荧光蛋白（GFP）标记的过表达ADAR异构体进行RNA免疫沉淀（RIP）测序显示，每个ADAR都与一组特定的差异表达基因相关，并且它们各自结合不同的RNA靶标。我们的结果显示与已知的编辑转录物有很好的重叠，建立了RIP-seq作为研究RNA编辑生物学的有效方法。
RNA editing by deamination of adenosine to inosine is an evolutionarily conserved process involved in many cellular pathways, from alternative splicing to miRNA targeting. In humans, it is carried out by no less than three major adenosine deaminases acting on RNA (ADARs): ADAR1-p150, ADAR1-p110, and ADAR2. However, the first two derive from alternative splicing, so that it is currently impossible to delete ADAR1-p110 without also knocking out ADAR1-p150 expression. Furthermore, the expression levels of ADARs varies wildly among cell types, and no study has systematically explored the effect of each of these isoforms on the cell transcriptome. In this study, RNA immunoprecipitation (RIP)-sequencing on overexpressed ADAR isoforms tagged with green fluorescent protein (GFP) shows that each ADAR is associated with a specific set of differentially expressed genes, and that they each bind to distinct set of RNA targets. Our results show a good overlap with known edited transcripts, establishing RIP-seq as a valid method for the investigation of RNA editing biology.