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Chemical RNA digestion enables robust RNA-binding site mapping at single amino acid resolution

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Title
Chemical RNA digestion enables robust RNA-binding site mapping at single amino acid resolution
Author(s)
Jong Woo Bae; S. Chul Kwon; Yongwoo Na; V. Narry Kim; Jong-Seo Kim
Subject
MASS-SPECTROMETRY, ; IDENTIFICATION, ; PHOSPHORYLATION, ; PROTEOME, ; PEPTIDE, ; DOMAINS
Publication Date
2020-07
Journal
NATURE STRUCTURAL & MOLECULAR BIOLOGY, v.27, no.7, pp.678 - 682
Publisher
NATURE PUBLISHING GROUP
Abstract
© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc. RNA-binding sites (RBSs) can be identified by liquid chromatography and tandem mass spectrometry analyses of the protein-RNA conjugates created by crosslinking, but RBS mapping remains highly challenging due to the complexity of the formed RNA adducts. Here, we introduce RBS-ID, a method that uses hydrofluoride to fully cleave RNA into mono-nucleosides, thereby minimizing the search space to drastically enhance coverage and to reach single amino acid resolution. Moreover, the simple mono-nucleoside adducts offer a confident and quantitative measure of direct RNA-protein interaction. Using RBS-ID, we profiled similar to 2,000 human RBSs and probedStreptococcus pyogenesCas9 to discover residues important for genome editing
URI
https://pr.ibs.re.kr/handle/8788114/7787
DOI
10.1038/s41594-020-0436-2
ISSN
1545-9993
Appears in Collections:
Center for RNA Research(RNA 연구단) > 1. Journal Papers (저널논문)
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