irCLASH reveals RNA substrates recognized by human ADARs
irCLASH reveals RNA substrates recognized by human ADARs
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irCLASH 揭示了人类 ADAR 识别的 RNA 底物
DOI：
10.1038/s41594-020-0398-4
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发表时间：
2020-03-23
影响因子：
16.8
通讯作者：
Zhang, Rui
中科院分区：
文献类型：
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作者：
Song, Yulong;Yang, Wenbing;Zhang, Rui
作用于RNA的腺苷脱氨酶（ADAR）在动物中将双链RNA（dsRNA）中的腺苷转化为肌苷。尽管其重要性，阿达尔RNA底物尚未在体内广泛定位。在这里，我们开发了irCLASH来映射人类ADAR识别的RNA底物，并揭示了决定其结合亲和力和编辑效率的特征。我们还观察到阿达尔底物内长程相互作用的优势，并分析ADAR 1和ADAR 2编辑底物之间的差异。此外，我们意外地发现，阿达尔蛋白在体内串联结合dsRNA底物，每个具有50-bp的足迹。使用RNA双链体识别ADARs作为前信使RNA结构的读出，我们揭示了前信使RNA和mRNA之间不同的高阶结构。我们的阿达尔底物的全转录组图谱和我们研究中观察到的RNA编辑的特征将有助于合理设计ADAR介导的RNA碱基编辑的指导RNA。
Adenosine deaminases acting on RNA (ADARs) convert adenosines to inosines in double-stranded RNA (dsRNA) in animals. Despite their importance, ADAR RNA substrates have not been mapped extensively in vivo. Here we develop irCLASH to map RNA substrates recognized by human ADARs and uncover features that determine their binding affinity and editing efficiency. We also observe a dominance of long-range interactions within ADAR substrates and analyze differences between ADAR1 and ADAR2 editing substrates. Moreover, we unexpectedly discovered that ADAR proteins bind dsRNA substrates tandemly in vivo, each with a 50-bp footprint. Using RNA duplexes recognized by ADARs as readout of pre-messenger RNA structures, we reveal distinct higher-order architectures between pre-messenger RNAs and mRNAs. Our transcriptome-wide atlas of ADAR substrates and the features governing RNA editing observed in our study will assist in the rational design of guide RNAs for ADAR-mediated RNA base editing.