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RING1 dictates GSDMD-mediated inflammatory response and host susceptibility to pathogen infection

Cell Death & Differentiation volume 32, pages 2066–2077 (2025)Cite this article

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Abstract

RING1 is an E3 ligase component of the polycomb repressive complex 1 (PRC1) with known roles in chromatin regulation and cellular processes such as apoptosis and autophagy. However, its involvement in inflammation and pyroptosis remains elusive. Here, we demonstrate that human RING1, not RING2, promotes K48-linked ubiquitination of Gasdermin D (GSDMD) and acts as a negative regulator of pyroptosis and bacterial infection. Indeed, we showed that loss of Ring1 increased S. typhimurium infectious load and mortality in vivo. Though RING1 deletion initially reduced M. tuberculosis (Mtb) infectious load in vivo, increased lung inflammation and impaired immune defense responses were later observed. Moreover, Ring1 knockout exacerbated acute sepsis induced by lipopolysaccharide (LPS) in vivo. Mechanistically, RING1 directly interacts with GSDMD and ubiquitinates the K51 and K168 sites of GSDMD for K48-linked proteasomal degradation, thereby inhibiting pyroptosis. Inhibition of RING1 E3 ligase activity by direct mutation or with the use of small molecule inhibitors increased GSDMD level and cell death during pyroptosis. Our findings reveal that RING1 dictates GSDMD-mediated inflammatory response and host susceptibility to pathogen infection, highlighting RING1 as a potential therapeutic target for combating infectious diseases.

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Fig. 1: RING1 deletion increases inflammatory response upon different pathogen infections.
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Fig. 2: Ring1 knockout mice are more susceptible to LPS-induced sepsis.
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Fig. 3: RING1 interacts with GSDMD.
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Fig. 4: GSDMD is ubiquitinated by RING1.
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Fig. 5: Ring1 knockout macrophages are more sensitive to pyroptosis.
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Fig. 6: Ring1 knockout upregulates GSDMD protein levels.
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Data availability

The RNA-seq data generated in this study have been deposited in the NCBI Sequence Read Archive (SRA). Raw data, including fastq files, have been uploaded under the accession numbers PRJNA1165469, PRJNA1165471, PRJNA1165473, PRJNA1165474, PRJNA1165475, PRJNA1165477, PRJNA1165479, and PRJNA1165480. All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the lead contact, J.L. (lijixi@fudan.edu.cn).

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (32161160323, 2018M641921) and the Shanghai Committee of Science and Technology (24490713600, 22YF1403400).

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Author notes

  1. These authors contributed equally: Yuanyuan Li, Wenqing Gao, Yuxin Qiu.

Authors and Affiliations

  1. State Key Laboratory of Genetics and Development of Complex Phenotypes, Department of Neurology, Huashan Hospital and School of Life Sciences, Fudan University, Shanghai, China

    Yuanyuan Li, Wenqing Gao, Yuxin Qiu, Jiasong Pan, Qingqing Guo, Xuehe Liu, Lu Geng, Yajie Shen, Yifan Deng & Jixi Li

  2. Centre for Immunology and Infection Control, School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD, Australia

    Wenqing Gao & Adi Idris

  3. Shanghai Institute of Infectious Diseases and Biosecurity & Shanghai Public Health Clinical Center, Fudan University, Shanghai, China

    Zhidong Hu & Xiaoyong Fan

  4. Division of Natural Science, Duke Kunshan University, Suzhou, China

    Suhua Li

  5. Shanghai Key Laboratory of Tuberculosis, Department of Microbiology and Immunology, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China

    Shanshan Liu, Hua Yang & Baoxue Ge

  6. Department of Chemistry, The Hong Kong University of Science and Technology, Hong Kong, China

    Jinqing Huang

  7. Department of Neurology, Huashan Hospital and Institute of Neurology, Fudan University, Shanghai, China

    Xiangjun Chen

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Contributions

JL conceived and designed the study. YL, WG, YQ, JP, QG, XL, LG, YS, YD, ZH, SL, SL, AI, JH, HY, BG, XF, and XC performed the experiments and analyzed the data. YL and JL analyzed the data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Jixi Li.

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The authors declare no competing interests.

Ethics

All animal experiments were performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals, with the approval of the Scientific Investigation Board of the School of Life Sciences, Fudan University (2020-JS-016).

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Li, Y., Gao, W., Qiu, Y. et al. RING1 dictates GSDMD-mediated inflammatory response and host susceptibility to pathogen infection. Cell Death Differ 32, 2066–2077 (2025). https://doi.org/10.1038/s41418-025-01527-2

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