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MAVS-loaded unanchored Lys63-linked polyubiquitin chains activate the RIG-I-MAVS signaling cascade

Cellular & Molecular Immunology volume 20, pages 1186–1202 (2023)Cite this article

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Abstract

The adaptor molecule MAVS forms prion-like aggregates to govern the RIG-I-like receptor (RLR) signaling cascade. Lys63 (K63)-linked polyubiquitination is critical for MAVS aggregation, yet the underlying mechanism and the corresponding E3 ligases and deubiquitinating enzymes (DUBs) remain elusive. Here, we found that the K63-linked polyubiquitin chains loaded on MAVS can be directly recognized by RIG-I to initiate RIG-I-mediated MAVS aggregation with the prerequisite of the CARDRIG-I-CARDMAVS interaction. Interestingly, many K63-linked polyubiquitin chains attach to MAVS via an unanchored linkage. We identified Ube2N as a major ubiquitin-conjugating enzyme for MAVS and revealed that Ube2N cooperates with the E3 ligase Riplet and TRIM31 to promote the unanchored K63-linked polyubiquitination of MAVS. In addition, we identified USP10 as a direct DUB that removes unanchored K63-linked polyubiquitin chains from MAVS. Consistently, USP10 attenuates RIG-I-mediated MAVS aggregation and the production of type I interferon. Mice with a deficiency in USP10 show more potent resistance to RNA virus infection. Our work proposes a previously unknown mechanism for the activation of the RLR signaling cascade triggered by MAVS-attached unanchored K63-linked polyubiquitin chains and establishes the DUB USP10 and the E2:E3 pair Ube2N-Riplet/TRIM31 as a specific regulatory system for the unanchored K63-linked ubiquitination and aggregation of MAVS upon viral infection.

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Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its supplementary information files or are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank Dr FaJian Hou (Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences) for the DDX58−/−, UBE2N−/− and MAVS−/− HEK293T cells. This work was supported by grants from the National Natural Science Foundation of China (31730026, 81930039, 32000633), National Key Research and Development Program (2021YFC2300603), Natural Science Foundation of Shandong Province (ZR2020QH136), China Postdoctoral Science Foundation (2020M682187), and Postdoctoral Innovation Project of Shandong Province (202002012).

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Authors and Affiliations

  1. Key Laboratory of Infection and Immunity of Shandong Province & Key Laboratory for Experimental Teratology of Ministry of Education, Shandong University, Jinan, 250012, Shandong, P.R. China

    Feng Liu, Wanxin Zhuang, Yuan Yang, Junqi Liu, Yi Zheng, Bingyu Liu, Wei Zhao & Chengjiang Gao

  2. Department of Immunology, School of Biomedical Sciences, Shandong University, Jinan, 250012, Shandong, P.R. China

    Feng Liu, Wanxin Zhuang, Yuan Yang, Junqi Liu, Yi Zheng, Bingyu Liu & Chengjiang Gao

  3. Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 201203, Shanghai, P.R. China

    Bin Song & Jie Zheng

  4. Department of Pathogenic Biology, School of Biomedical Sciences, Shandong University, Jinan, 250012, Shandong, P.R. China

    Wei Zhao

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  1. Feng Liu
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Contributions

CG conceived the research; CG and FL designed the experiments; FL performed the research; WZ, BS, YY, and JL provided reagents and participated in the experiments; JZ revised the paper and participated in discussions; YZ, BL and WZ participated in discussions; CG and FL analyzed the data; CG and FL wrote the paper.

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Correspondence to Chengjiang Gao.

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Liu, F., Zhuang, W., Song, B. et al. MAVS-loaded unanchored Lys63-linked polyubiquitin chains activate the RIG-I-MAVS signaling cascade. Cell Mol Immunol 20, 1186–1202 (2023). https://doi.org/10.1038/s41423-023-01065-2

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