Wu Tong, Cheng Anthony Youzhi, Zhang Yuexiu, Xu Jiayu, Wu Jinjun, Wen Li, Li Xiao, Liu Bei, Dou Xiaoyang, Wang Pingluan, Zhang Linda, Fei Jingyi, Li Jianrong, Ouyang Zhengqing, He Chuan
Department of Chemistry, University of Chicago, Chicago, IL, USA.
Howard Hughes Medical Institute, Chicago, IL, USA.
Nat Biotechnol. 2024 Dec;42(12):1909-1920. doi: 10.1038/s41587-023-02109-8. Epub 2024 Jan 18.
RNA fate and function are affected by their structures and interactomes. However, how RNA and RNA-binding proteins (RBPs) assemble into higher-order structures and how RNA molecules may interact with each other to facilitate functions remain largely unknown. Here we present KARR-seq, which uses N-kethoxal labeling and multifunctional chemical crosslinkers to covalently trap and determine RNA-RNA interactions and higher-order RNA structures inside cells, independent of local protein binding to RNA. KARR-seq depicts higher-order RNA structure and detects widespread intermolecular RNA-RNA interactions with high sensitivity and accuracy. Using KARR-seq, we show that translation represses mRNA compaction under native and stress conditions. We determined the higher-order RNA structures of respiratory syncytial virus (RSV) and vesicular stomatitis virus (VSV) and identified RNA-RNA interactions between the viruses and the host RNAs that potentially regulate viral replication.
RNA的命运和功能受其结构和相互作用组的影响。然而,RNA与RNA结合蛋白(RBP)如何组装成高阶结构,以及RNA分子如何相互作用以促进功能,在很大程度上仍然未知。在此,我们介绍了KARR-seq,它利用N-乙二醛标记和多功能化学交联剂,在细胞内共价捕获并确定RNA-RNA相互作用和高阶RNA结构,而不依赖于局部蛋白质与RNA的结合。KARR-seq描绘了高阶RNA结构,并以高灵敏度和准确性检测广泛的分子间RNA-RNA相互作用。利用KARR-seq,我们表明在天然和应激条件下,翻译会抑制mRNA压缩。我们确定了呼吸道合胞病毒(RSV)和水疱性口炎病毒(VSV)的高阶RNA结构,并鉴定了病毒与宿主RNA之间潜在调节病毒复制的RNA-RNA相互作用。
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