Laboratory of Genetics and Disorders, Key Laboratory of Molecular Medicine and Biotherapy, Aerospace Center Hospital, School of Life Science, Beijing Institute of Technology, Beijing, People's Republic of China.
Advanced Technology Research Institute, Beijing Institute of Technology, Jinan, People's Republic of China.
Wiley Interdiscip Rev RNA. 2024 May-Jun;15(3):e1863. doi: 10.1002/wrna.1863.
RNA-binding proteins (RBPs) are powerful and versatile regulators in living creatures, playing fundamental roles in organismal development, metabolism, and various diseases by the regulation of gene expression at multiple levels. The requirements of deep research on RBP function have promoted the rapid development of RBP-RNA interplay detection methods. Recently, the detection method of fusing RNA modification enzymes (RME) with RBP of interest has become a hot topic. Here, we reviewed RNA modification enzymes in adenosine deaminases that act on RNA (ADAR), terminal nucleotidyl transferase (TENT), and activation-induced cytosine deaminase/ApoB mRNA editing enzyme catalytic polypeptide-like (AID/APOBEC) protein family, regarding the biological function, biochemical activity, and substrate specificity originated from enzyme selves, their domains and partner proteins. In addition, we discussed the RME activity screening system, and the RME mutations with engineered enzyme activity. Furthermore, we provided a systematic overview of the basic principles, advantages, disadvantages, and applications of the RME-based and cross-linking and immunopurification (CLIP)-based RBP target profiling strategies, including targets of RNA-binding proteins identified by editing (TRIBE), RNA tagging, surveying targets by APOBEC-mediated profiling (STAMP), CLIP-seq, and their derivative technology. This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein-RNA Recognition RNA Processing > RNA Editing and Modification.
RNA 结合蛋白 (RBPs) 是生物体内强大而多功能的调控因子，通过在多个层面上调节基因表达，在生物个体发育、代谢和各种疾病中发挥着基本作用。对 RBP 功能进行深入研究的需求促进了 RBP-RNA 相互作用检测方法的快速发展。最近，将 RNA 修饰酶 (RME) 与感兴趣的 RBP 融合的检测方法成为一个热门话题。在这里，我们综述了腺苷脱氨酶作用于 RNA（ADAR）、末端核苷酸转移酶 (TENT) 和激活诱导胞嘧啶脱氨酶/载脂蛋白 B mRNA 编辑酶催化多肽样 (AID/APOBEC) 蛋白家族中的 RNA 修饰酶的生物学功能、生化活性和底物特异性，这些酶源于自身的酶学活性、结构域和伴侣蛋白。此外，我们还讨论了 RME 活性筛选系统以及具有工程酶活性的 RME 突变。此外，我们还系统地概述了基于 RME 和交联免疫沉淀 (CLIP) 的 RBP 靶标分析策略的基本原理、优缺点及其应用，包括通过编辑识别的 RNA 结合蛋白靶标 (TRIBE)、RNA 标记、APOBEC 介导的靶标分析 (STAMP)、CLIP-seq 及其衍生技术。本文属于以下类别：RNA 与蛋白质和其他分子的相互作用 > 蛋白质-RNA 识别 RNA 加工 > RNA 编辑和修饰。
