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KARR-seq技术揭示了细胞中的高阶RNA结构以及RNA-RNA相互作用。

KARR-seq reveals cellular higher-order RNA structures and RNA-RNA interactions.

作者信息

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

机构信息

Department of Chemistry, University of Chicago, Chicago, IL, USA.

Howard Hughes Medical Institute, Chicago, IL, USA.

出版信息

Nat Biotechnol. 2024 Dec;42(12):1909-1920. doi: 10.1038/s41587-023-02109-8. Epub 2024 Jan 18.


DOI:10.1038/s41587-023-02109-8
PMID:38238480
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11255127/
Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/46ead1831d64/41587_2023_2109_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/32b1ec20436c/41587_2023_2109_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/e9dbb19fb097/41587_2023_2109_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/13833ade1d24/41587_2023_2109_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/7eed294ace69/41587_2023_2109_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/46ead1831d64/41587_2023_2109_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/32b1ec20436c/41587_2023_2109_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/e9dbb19fb097/41587_2023_2109_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/13833ade1d24/41587_2023_2109_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/7eed294ace69/41587_2023_2109_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55b4/11631761/46ead1831d64/41587_2023_2109_Fig5_HTML.jpg

相似文献

[1]
KARR-seq reveals cellular higher-order RNA structures and RNA-RNA interactions.

Nat Biotechnol. 2024-12

[2]
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[3]
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[4]
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[5]
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J Virol. 2010-9-15

[6]
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Semin Cell Dev Biol. 2021-3

[7]
Discontinuous L-binding motifs in the transactivation domain of the vesicular stomatitis virus P protein are required for terminal transcription initiation by the L protein.

J Virol. 2023-8-31

[8]
Diverse roles of host RNA binding proteins in RNA virus replication.

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[9]
higher-order oligomeric assembly of the respiratory syncytial virus M2-1 protein with longer RNAs.

J Virol. 2024-8-20

[10]
Targeting RNA decay with 2',5' oligoadenylate-antisense in respiratory syncytial virus-infected cells.

Proc Natl Acad Sci U S A. 1997-3-4

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[3]
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[4]
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[5]
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[6]
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[7]
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[10]
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本文引用的文献

[1]
A computationally-enhanced hiCLIP atlas reveals Staufen1-RNA binding features and links 3' UTR structure to RNA metabolism.

Nucleic Acids Res. 2023-5-8

[2]
GSEApy: a comprehensive package for performing gene set enrichment analysis in Python.

Bioinformatics. 2023-1-1

[3]
Structural basis of human telomerase recruitment by TPP1-POT1.

Science. 2022-3-11

[4]
RNA promotes the formation of spatial compartments in the nucleus.

Cell. 2021-11-11

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Comprehensive mapping of SARS-CoV-2 interactions in vivo reveals functional virus-host interactions.

Nat Commun. 2021-8-25

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Direct Mapping of Higher-Order RNA Interactions by SHAPE-JuMP.

Biochemistry. 2021-6-29

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Optimized photochemistry enables efficient analysis of dynamic RNA structuromes and interactomes in genetic and infectious diseases.

Nat Commun. 2021-4-20

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Heterogeneous Dynamics of Protein-RNA Interactions across Transcriptome-Derived Messenger RNA Populations.

J Am Chem Soc. 2020-12-23

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The Short- and Long-Range RNA-RNA Interactome of SARS-CoV-2.

Mol Cell. 2020-12-17

[10]
Direct DNA crosslinking with CAP-C uncovers transcription-dependent chromatin organization at high resolution.

Nat Biotechnol. 2021-2

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