期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 61, 期 27, 页码 -出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202202008
关键词
Crosslinking; Photocatalysis; RNA-Binding Protein; RNA-Protein Interactome; SARS-CoV-2
资金
- Ministry of Science and Technology [2018YFA0507600, 2016YFA0501500, 2017YFA0503600]
- National Natural Science Foundation of China [22007001, 32088101, 21727806, 91753131, 91753000, 91957101, 22077004]
- Beijing Natural Science Foundation [Z200010]
- Li Ge-Zhao Ning Life Science Junior Research Fellowship
- Bayer Investigator Award
The interactions between RNA and proteins are crucial for cellular processes. This study developed a photocatalytic crosslinking strategy combined with mass spectrometry and RNA sequencing to study the composition and dynamics of protein-RNA interactions. The approach identified numerous RNA binding proteins in human cells and investigated the dynamic changes of RBPs in macrophage cells and their interactions with SARS-CoV-2 RNA.
The dynamic interactions between RNAs and proteins play crucial roles in regulating diverse cellular processes. Proteome-wide characterization of these interactions in their native cellular context remains desirable but challenging. Herein, we developed a photocatalytic crosslinking (PhotoCAX) strategy coupled with mass spectrometry (PhotoCAX-MS) and RNA sequencing (PhotoCAX-seq) for the study of the composition and dynamics of protein-RNA interactions. By integrating the blue light-triggered photocatalyst with a dual-functional RNA-protein crosslinker (RP-linker) and the phase separation-based enrichment strategy, PhotoCAX-MS revealed a total of 2044 RBPs in human HEK293 cells. We further employed PhotoCAX to investigate the dynamic change of RBPome in macrophage cells upon LPS-stimulation, as well as the identification of RBPs interacting directly with the 5 ' untranslated regions of SARS-CoV-2 RNA.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据