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TRIM21 regulates pyroptotic cell death by promoting Gasdermin D oligomerization

Cell Death & Differentiation volume 29, pages 439–450 (2022)Cite this article

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Abstract

Gasdermin-D (GSDMD), the executioner of pyroptotic cell death when it is cleaved by inflammatory caspases, plays a crucial role in host defense and the response to danger signals. So far, there are no known mechanisms, other than cleavage, for regulating GSDMD. Here, we show that tripartite motif protein TRIM21 acts as a positive regulator of GSDMD-dependent pyroptosis. TRIM21 interacted with GSDMD via its PRY-SPRY domain, maintaining GSDMD stable expression in resting cells yet inducing the N-terminus of GSDMD (GSDMD-N) aggregation during pyroptosis. TRIM21-deficient cells displayed a reduced cell death in response to NLRP3 or NLRC4 inflammasome activation. Genetic ablation of TRIM21 in mice conferred protection from LPS-induced inflammation and dextran sulfate sodium-induced colitis. Therefore, TRIM21 plays an essential role in GSDMD-mediated pyroptosis and may be a viable target for controlling and treating inflammation-associated diseases.

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Fig. 1: GSDMD directly interacts with TRIM21.
Fig. 2: TRIM21 binds GSDMD via its PRY-SPRY domain.
Fig. 3: TRIM21 enhances the stability of GSDMD independent of its E3 ligase activity.
Fig. 4: Depletion of TRIM21 alleviates pyroptosis.
Fig. 5: Depletion of TRIM21 reduces GSDMD-N upon inflammatory stimulation.
Fig. 6: The PRY-SPRY domain of TRIM21 promotes GSDMD-N oligomerization and cell death.
Fig. 7: TRIM21 deficiency ameliorates LPS-induced inflammation and DSS-induced colitis.

Data availability

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 corresponding author.

References

  1. Galluzzi L, Vitale I, Aaronson SA, Abrams JM, Adam D, Agostinis P, et al. Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ. 2018;25:486–541.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Kovacs SB, Miao EA. Gasdermins: effectors of pyroptosis. Trends Cell Biol. 2017;27:673–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Li J, McQuade T, Siemer AB, Napetschnig J, Moriwaki K, Hsiao YS, et al. The RIP1/RIP3 necrosome forms a functional amyloid signaling complex required for programmed necrosis. Cell. 2012;150:339–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Mompeán M, Li W, Li J, Laage S, Siemer AB, Bozkurt G, et al. The structure of the necrosome RIPK1-RIPK3 core, a human hetero-amyloid signaling complex. Cell. 2018;173:1244–53.e1210.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Green DR. The coming decade of cell death research: five riddles. Cell. 2019;177:1094–107.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kayagaki N, Stowe IB, Lee BL, O’Rourke K, Anderson K, Warming S, et al. Caspase-11 cleaves gasdermin D for non-canonical inflammasome signalling. Nature. 2015;526:666–71.

    Article  CAS  PubMed  Google Scholar 

  7. Shi J, Zhao Y, Wang K, Shi X, Wang Y, Huang H, et al. Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death. Nature. 2015;526:660–5.

    Article  CAS  PubMed  Google Scholar 

  8. Liu X, Zhang Z, Ruan J, Pan Y, Magupalli VG, Wu H, et al. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature. 2016;535:153–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Martinon F, Tschopp J. Inflammatory caspases: linking an intracellular innate immune system to autoinflammatory diseases. Cell. 2004;117:561–74.

    Article  CAS  PubMed  Google Scholar 

  10. Man SM, Kanneganti TD, Gasdermin D. the long-awaited executioner of pyroptosis. Cell Res. 2015;25:1183–4.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Ruan J, Xia S, Liu X, Lieberman J, Wu H. Cryo-EM structure of the gasdermin A3 membrane pore. Nature. 2018;557:62–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Chen X, He WT, Hu L, Li J, Fang Y, Wang X, et al. Pyroptosis is driven by non-selective gasdermin-D pore and its morphology is different from MLKL channel-mediated necroptosis. Cell Res. 2016;26:1007–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Wright JA, Bryant CE. The killer protein Gasdermin D. Cell Death Differ. 2016;23:1897–8.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Ding J, Wang K, Liu W, She Y, Sun Q, Shi J, et al. Pore-forming activity and structural autoinhibition of the gasdermin family. Nature. 2016;535:111–6.

    Article  CAS  PubMed  Google Scholar 

  15. Kuang S, Zheng J, Yang H, Li S, Duan S, Shen Y, et al. Structure insight of GSDMD reveals the basis of GSDMD autoinhibition in cell pyroptosis. Proc Natl Acad Sci USA. 2017;114:10642–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Liu Z, Wang C, Yang J, Zhou B, Yang R, Ramachandran R, et al. Crystal structures of the full-length murine and human gasdermin D reveal mechanisms of autoinhibition, lipid binding, and oligomerization. Immunity. 2019;51:43–9.e44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Orning P, Weng D, Starheim K, Ratner D, Best Z, Lee B, et al. Pathogen blockade of TAK1 triggers caspase-8-dependent cleavage of gasdermin D and cell death. Science. 2018;362:1064–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Evavold CL, Ruan J, Tan Y, Xia S, Wu H, Kagan JC. The pore-forming protein gasdermin D regulates interleukin-1 secretion from living macrophages. Immunity. 2018;48:35–44.e36.

    Article  CAS  PubMed  Google Scholar 

  19. Ruhl S, Shkarina K, Demarco B, Heilig R, Santos JC, Broz P. ESCRT-dependent membrane repair negatively regulates pyroptosis downstream of GSDMD activation. Science. 2018;362:956–60.

    Article  PubMed  Google Scholar 

  20. Hatakeyama S. TRIM family proteins: roles in autophagy, immunity, and carcinogenesis. Trends Biochem Sci. 2017;42:297–311.

    Article  CAS  PubMed  Google Scholar 

  21. Oke V, Wahren-Herlenius M. The immunobiology of Ro52 (TRIM21) in autoimmunity: a critical review. J Autoimmun. 2012;39:77–82.

    Article  CAS  PubMed  Google Scholar 

  22. Xue B, Li H, Guo M, Wang J, Xu Y, Zou X, et al. TRIM21 promotes innate immune response to RNA viral infection through Lys27-linked polyubiquitination of MAVS. J Virol. 2018;92.

  23. Pan JA, Sun Y, Jiang YP, Bott AJ, Jaber N, Dou Z, et al. TRIM21 ubiquitylates SQSTM1/p62 and suppresses protein sequestration to regulate redox homeostasis. Mol Cell. 2016;61:720–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Samir P, Kesavardhana S, Patmore DM, Gingras S, Malireddi RKS, Karki R, et al. DDX3X acts as a live-or-die checkpoint in stressed cells by regulating NLRP3 inflammasome. Nature. 2019;573:590–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Wyllie DH, Kiss-Toth E, Visintin A, Smith SC, Boussouf S, Segal DM, et al. Evidence for an accessory protein function for Toll-like receptor 1 in anti-bacterial responses. J Immunol. 2000;165:7125–32.

    Article  CAS  PubMed  Google Scholar 

  26. Rogers C, Erkes DA, Nardone A, Aplin AE, Fernandes-Alnemri T, Alnemri ES. Gasdermin pores permeabilize mitochondria to augment caspase-3 activation during apoptosis and inflammasome activation. Nat Commun. 2019;10:1689.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Qiu S, Liu J, Xing F. ‘Hints’ in the killer protein gasdermin D: unveiling the secrets of gasdermins driving cell death. Cell Death Differ. 2017;24:588–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Kayagaki N, Warming S, Lamkanfi M, Vande Walle L, Louie S, Dong J, et al. Non-canonical inflammasome activation targets caspase-11. Nature. 2011;479:117–21.

    Article  CAS  PubMed  Google Scholar 

  29. Bozza FA, Salluh JI, Japiassu AM, Soares M, Assis EF, Gomes RN, et al. Cytokine profiles as markers of disease severity in sepsis: a multiplex analysis. Crit Care. 2007;11:R49.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Aglietti RA, Dueber EC. Recent insights into the molecular mechanisms underlying pyroptosis and gasdermin family functions. Trends Immunol. 2017;38:261–71.

    Article  CAS  PubMed  Google Scholar 

  31. Wirtz S, Neufert C, Weigmann B, Neurath MF. Chemically induced mouse models of intestinal inflammation. Nat Protoc. 2007;2:541–6.

    Article  CAS  PubMed  Google Scholar 

  32. Shi J, Gao W, Shao F. Pyroptosis: gasdermin-mediated programmed necrotic cell death. Trends Biochem Sci. 2017;42:245–54.

    Article  CAS  PubMed  Google Scholar 

  33. Rhodes DA, Isenberg DA. TRIM21 and the function of antibodies inside cells. Trends Immunol. 2017;38:916–26.

    Article  CAS  PubMed  Google Scholar 

  34. Jiang X, Chen ZJ. The role of ubiquitylation in immune defence and pathogen evasion. Nat Rev Immunol. 2011;12:35–48.

    Article  PubMed  Google Scholar 

  35. Cadena C, Ahmad S, Xavier A, Willemsen J, Park S, Park JW, et al. Ubiquitin-dependent and -independent roles of E3 ligase RIPLET in innate immunity. Cell. 2019;177:1187–200.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Gack MU, Shin YC, Joo CH, Urano T, Liang C, Sun L, et al. TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity. Nature. 2007;446:916–20.

    Article  CAS  PubMed  Google Scholar 

  37. Yan J, Li Q, Mao AP, Hu MM, Shu HB. TRIM4 modulates type I interferon induction and cellular antiviral response by targeting RIG-I for K63-linked ubiquitination. J Mol Cell Biol. 2014;6:154–63.

    Article  CAS  PubMed  Google Scholar 

  38. Liu B, Zhang M, Chu H, Zhang H, Wu H, Song G, et al. The ubiquitin E3 ligase TRIM31 promotes aggregation and activation of the signaling adaptor MAVS through Lys63-linked polyubiquitination. Nat Immunol. 2017;18:214–24.

    Article  CAS  PubMed  Google Scholar 

  39. Seo GJ, Kim C, Shin WJ, Sklan EH, Eoh H, Jung JU. TRIM56-mediated monoubiquitination of cGAS for cytosolic DNA sensing. Nat Commun. 2018;9:613.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Yang L, Jin L, Ke Y, Fan X, Zhang T, Zhang C, et al. E3 Ligase Trim21 ubiquitylates and stabilizes keratin 17 to induce STAT3 activation in psoriasis. J Investig Dermatol. 2018;138:2568–77.

    Article  CAS  PubMed  Google Scholar 

  41. Sanchez JG, Sparrer KMJ, Chiang C, Reis RA, Chiang JJ, Zurenski MA, et al. TRIM25 binds RNA to modulate cellular anti-viral defense. J Mol Biol. 2018;430:5280–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Yap MW, Nisole S, Stoye JP. A single amino acid change in the SPRY domain of human Trim5alpha leads to HIV-1 restriction. Curr Biol. 2005;15:73–8.

    Article  CAS  PubMed  Google Scholar 

  43. Pan JA, Sun Y, Jiang YP, Bott AJ, Jaber N, Dou Z, et al. TRIM21 ubiquitylates SQSTM1/p62 and suppresses protein sequestration to regulate redox homeostasis. Mol Cell. 2016;62:149–51.

    Article  CAS  PubMed  Google Scholar 

  44. James LC, Keeble AH, Khan Z, Rhodes DA, Trowsdale J. Structural basis for PRYSPRY-mediated tripartite motif (TRIM) protein function. Proc Natl Acad Sci USA. 2007;104:6200–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Wang Y, Gao W, Shi X, Ding J, Liu W, He H, et al. Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin. Nature. 2017;547:99–103.

    Article  CAS  PubMed  Google Scholar 

  46. Zhou G, Wu W, Yu L, Yu T, Yang W, Wang P, et al. Tripartite motif-containing (TRIM) 21 negatively regulates intestinal mucosal inflammation through inhibiting TH1/TH17 cell differentiation in patients with inflammatory bowel diseases. J Allergy Clin Immunol. 2018;142:1218–28.e1212.

    Article  CAS  PubMed  Google Scholar 

  47. Assouvie A, Daley-Bauer LP, Rousselet G. Growing murine bone marrow-derived macrophages. Methods Mol Biol. 2018;1784:29–33.

    Article  CAS  PubMed  Google Scholar 

  48. Pineda-Torra I, Gage M, de Juan A, Pello OM. Isolation, culture, and polarization of murine bone marrow-derived and peritoneal macrophages. Methods Mol Biol. 2015;1339:101–9.

    Article  CAS  PubMed  Google Scholar 

  49. Sacks D, Baxter B, Campbell BCV, Carpenter JS, Cognard C, Dippel D, et al. Multisociety consensus quality improvement revised consensus statement for endovascular therapy of acute ischemic stroke. Int J Stroke. 2018;13:612–32.

    PubMed  Google Scholar 

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Acknowledgements

We thank Dr. H. Saiyin for the help in mouse tissue IHC experiments. Gsdmd−/− HeLa cells and Gsdmd−/− iBMDMs are kindly provided by Dr. F. Shao (National Institute of Biological Sciences, China). Gsdmd−/− mice are kindly provided by Dr. Z. Lin (Nanjing University, China). WT pathogenic Salmonella typhimurium (SL14028s) is kindly provided by Dr. Y. Yao (Shanghai Jiaotong University, China).

Funding

This work was supported by grants from the National Natural Science Foundation of China (82071782, 31670878), the National Key Research and Development Project of China (2016YFA0500600), and the Shanghai Committee of Science and Technology (20XD1400800).

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

Authors and Affiliations

  1. Department of Neurology, Huashan Hospital and Institute of Neurology, State Key Laboratory of Genetic Engineering, School of Life Sciences, MOE Engineering Research Center of Gene Technology, Shanghai Engineering Research Center of Industrial Microorganisms, Fudan University, Shanghai, China

    Wenqing Gao, Yuanyuan Li, Xuehe Liu, Sen Wang, Pucheng Mei, Zijun Chen & Jixi Li

  2. Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China

    Kewei Liu & Yuping Lai

  3. Division of Natural Science, Duke Kunshan University, Suzhou, Jiangsu, China

    Suhua Li

  4. Key Laboratory of Marine Ecosystem Dynamics, Ministry of Natural Resources & Second Institute of Oceanography, Ministry of Natural Resources, Hangzhou, China

    Xue-Wei Xu

  5. State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, China

    Jianhua Gan, Jiaxue Wu, Chaoneng Ji & Chen Ding

  6. Program in Cellular and Molecular Medicine, Boston Children’s Hospital and Department of Pediatrics, Harvard Medical School, Boston, MA, USA

    Xing Liu & Judy Lieberman

  7. Department of Medical Biophysics, University of Toronto, and Princess Margaret Cancer Center, University Health Network, Toronto, ON, Canada

    Housheng Hansen He

  8. Program in Cellular and Molecular Medicine, Boston Children’s Hospital and Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA

    Hao Wu

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

    Xiangjun Chen

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Contributions

JL, XC, and WG conceived and designed the study. WG, YL, XL, SW, PM, ZC, KL, SL, XX, JG, JW, CJ, CD, XL, YL, HH, JL, and HW performed the experiments and analyzed the data. JL and WG analyzed the data and wrote the paper. All authors discussed the results and commented on the paper.

Corresponding authors

Correspondence to Xiangjun Chen or Jixi Li.

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The authors declare no competing interests.

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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 School of Life Sciences, Fudan University (2019-JS-011).

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Gao, W., Li, Y., Liu, X. et al. TRIM21 regulates pyroptotic cell death by promoting Gasdermin D oligomerization. Cell Death Differ 29, 439–450 (2022). https://doi.org/10.1038/s41418-021-00867-z

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