Revealing the hidden RBP–RNA interactions with RNA modification enzyme‐based strategies

Abstract

AbstractRNA‐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