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USP3 deubiquitinates and stabilizes the adapter protein ASC to regulate inflammasome activation

Abstract

Inflammasomes are essential components of the innate immune system and its defense against infections, whereas the dysregulation of inflammasome activation has a detrimental effect on human health. The activation of inflammasomes is subjected to tight regulation to maintain immune homeostasis, yet the underlying mechanism remains elusive. Here, we identify USP3 as a direct deubiquitinating enzyme (DUB) for ASC, the central adapter mediating the assembly and activation of most inflammasomes. USP3 removes the K48-linked ubiquitination on ASC and strengthens its stability by blocking proteasomal degradation. Additionally, USP3 promotes inflammasome activation, and this function was confirmed in mouse models of aluminum (Alum)-induced peritonitis, F. novicida infection and flagellin-induced pneumonia in vivo. Our work unveils that USP3 functions as a key regulator of ASC ubiquitination and maintains the physiological role of ASC in mediating inflammasome activation, and we propose a new mechanism by which the ubiquitination of ASC regulates inflammasome activation.

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References

  1. Broz P, Dixit VM. Inflammasomes: mechanism of assembly, regulation and signalling. Nat Rev Immunol. 2016;16:407–20.

    Article  CAS  Google Scholar 

  2. Swanson KV, Deng M, Ting JP. The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol. 2019;19:477–89.

    Article  CAS  Google Scholar 

  3. Schroder K, Tschopp J. The inflammasomes. Cell. 2010;140:821–32.

    Article  CAS  Google Scholar 

  4. Guo H, Callaway JB, Ting JP. Inflammasomes: mechanism of action, role in disease, and therapeutics. Nat Med. 2015;21:677–87.

    Article  Google Scholar 

  5. Milner MT, Maddugoda M, Gotz J, Burgener SS, Schroder K. The NLRP3 inflammasome triggers sterile neuroinflammation and Alzheimer’s disease. Curr Opin Immunol. 2021;68:116–24.

    Article  CAS  Google Scholar 

  6. Davis BK, Wen H, Ting JP. The inflammasome NLRs in immunity, inflammation, and associated diseases. Annu Rev Immunol. 2011;29:707–35.

    Article  CAS  Google Scholar 

  7. Ge Q, Chen X, Zhao Y, Mu H, Zhang J. Modulatory mechanisms of NLRP3: Potential roles in inflammasome activation. Life Sci. 2021;267:118918.

    Article  CAS  Google Scholar 

  8. Lopez-Castejon G. Control of the inflammasome by the ubiquitin system. Febs j. 2020;287:11–26.

    Article  CAS  Google Scholar 

  9. Shim DW, Lee KH. Posttranslational Regulation of the NLR Family Pyrin Domain-Containing 3 Inflammasome. Front Immunol. 2018;9:1054.

    Article  Google Scholar 

  10. Popovic D, Vucic D, Dikic I. Ubiquitination in disease pathogenesis and treatment. Nat Med. 2014;20:1242–53.

    Article  CAS  Google Scholar 

  11. Humphries F, Bergin R, Jackson R, Delagic N, Wang B, Yang S, et al. The E3 ubiquitin ligase Pellino2 mediates priming of the NLRP3 inflammasome. Nat Commun. 2018;9:1560.

    Article  Google Scholar 

  12. Song H, Liu B, Huai W, Yu Z, Wang W, Zhao J, et al. The E3 ubiquitin ligase TRIM31 attenuates NLRP3 inflammasome activation by promoting proteasomal degradation of NLRP3. Nat Commun. 2016;7:13727.

    Article  CAS  Google Scholar 

  13. Py BF, Kim MS, Vakifahmetoglu-Norberg H, Yuan J. Deubiquitination of NLRP3 by BRCC3 critically regulates inflammasome activity. Mol Cell. 2013;49:331–8.

    Article  CAS  Google Scholar 

  14. Palazon-Riquelme P, Worboys JD, Green J, Valera A, Martin-Sanchez F, Pellegrini C, et al. USP7 and USP47 deubiquitinases regulate NLRP3 inflammasome activation. EMBO Rep. 2018;19:e44766.

  15. Hoss F, Rodriguez-Alcazar JF, Latz E. Assembly and regulation of ASC specks. Cell Mol Life Sci. 2017;74:1211–29.

    Article  CAS  Google Scholar 

  16. Dick MS, Sborgi L, Ruhl S, Hiller S, Broz P. ASC filament formation serves as a signal amplification mechanism for inflammasomes. Nat Commun. 2016;7:11929.

    Article  CAS  Google Scholar 

  17. Rodgers MA, Bowman JW, Fujita H, Orazio N, Shi M, Liang Q, et al. The linear ubiquitin assembly complex (LUBAC) is essential for NLRP3 inflammasome activation. J Exp Med. 2014;211:1333–47.

    Article  CAS  Google Scholar 

  18. Shi CS, Shenderov K, Huang NN, Kabat J, Abu-Asab M, Fitzgerald KA, et al. Activation of autophagy by inflammatory signals limits IL-1beta production by targeting ubiquitinated inflammasomes for destruction. Nat Immunol. 2012;13:255–63.

    Article  CAS  Google Scholar 

  19. Guan K, Wei C, Zheng Z, Song T, Wu F, Zhang Y, et al. MAVS Promotes Inflammasome Activation by Targeting ASC for K63-Linked Ubiquitination via the E3 Ligase TRAF3. J Immunol. 2015;194:4880–90.

    Article  CAS  Google Scholar 

  20. Siu KL, Yuen KS, Castaño-Rodriguez C, Ye ZW, Yeung ML, Fung SY, et al. Severe acute respiratory syndrome coronavirus ORF3a protein activates the NLRP3 inflammasome by promoting TRAF3-dependent ubiquitination of ASC. Faseb j. 2019;33:8865–77.

    Article  CAS  Google Scholar 

  21. Zhang L, Ko CJ, Li Y, Jie Z, Zhu L, Zhou X, et al. Peli1 facilitates NLRP3 inflammasome activation by mediating ASC ubiquitination. Cell Rep. 2021;37:109904.

    Article  CAS  Google Scholar 

  22. Lee JY, Seo D, You J, Chung S, Park JS, Lee JH, et al. The deubiquitinating enzyme, ubiquitin-specific peptidase 50, regulates inflammasome activation by targeting the ASC adaptor protein. FEBS Lett. 2017;591:479–90.

    Article  CAS  Google Scholar 

  23. Cheng YC, Shieh SY. Deubiquitinating enzyme USP3 controls CHK1 chromatin association and activation. Proc Natl Acad Sci USA. 2018;115:5546–51.

    Article  CAS  Google Scholar 

  24. Grice GL, Nathan JA. The recognition of ubiquitinated proteins by the proteasome. Cell Mol Life Sci. 2016;73:3497–506.

    Article  CAS  Google Scholar 

  25. Cai X, Xu H, Chen ZJ. Prion-Like Polymerization in Immunity and Inflammation. Cold Spring Harb Perspect Biol. 2017;9:a023580.

  26. Pelegrin P, Barroso-Gutierrez C, Surprenant A. P2X7 receptor differentially couples to distinct release pathways for IL-1beta in mouse macrophage. J Immunol. 2008;180:7147–57.

    Article  CAS  Google Scholar 

  27. Eisenbarth SC, Colegio OR, O’Connor W, Sutterwala FS, Flavell RA. Crucial role for the Nalp3 inflammasome in the immunostimulatory properties of aluminium adjuvants. Nature. 2008;453:1122–6.

    Article  CAS  Google Scholar 

  28. Sokolovska A, Hem SL, HogenEsch H. Activation of dendritic cells and induction of CD4(+) T cell differentiation by aluminum-containing adjuvants. Vaccine. 2007;25:4575–85.

    Article  CAS  Google Scholar 

  29. Guarda G, Braun M, Staehli F, Tardivel A, Mattmann C, Forster I, et al. Type I interferon inhibits interleukin-1 production and inflammasome activation. Immunity. 2011;34:213–23.

    Article  CAS  Google Scholar 

  30. Guo Y, Li L, Xu T, Guo X, Wang C, Li Y, et al. HUWE1 mediates inflammasome activation and promotes host defense against bacterial infection. J Clin Investig. 2020;130:6301–16.

    Article  CAS  Google Scholar 

  31. Xue Y, Enosi Tuipulotu D, Tan WH, Kay C, Man SM. Emerging Activators and Regulators of Inflammasomes and Pyroptosis. Trends Immunol. 2019;40:1035–52.

    Article  CAS  Google Scholar 

  32. Cui J, Song Y, Li Y, Zhu Q, Tan P, Qin Y, et al. USP3 inhibits type I interferon signaling by deubiquitinating RIG-I-like receptors. Cell Res. 2014;24:400–16.

    Article  CAS  Google Scholar 

  33. Fu S, Shao S, Wang L, Liu H, Hou H, Wang Y, et al. USP3 stabilizes p53 protein through its deubiquitinase activity. Biochem Biophys Res Commun. 2017;492:178–83.

    Article  CAS  Google Scholar 

  34. Nicassio F, Corrado N, Vissers JH, Areces LB, Bergink S, Marteijn JA, et al. Human USP3 is a chromatin modifier required for S phase progression and genome stability. Curr Biol. 2007;17:1972–7.

    Article  CAS  Google Scholar 

  35. Shi K, Zhang JZ, Yang L, Li NN, Yue Y, Du XH, et al. Protein deubiquitylase USP3 stabilizes Aurora A to promote proliferation and metastasis of esophageal squamous cell carcinoma. BMC Cancer. 2021;21:1196.

    Article  CAS  Google Scholar 

  36. 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  Google Scholar 

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Acknowledgements

This work was supported by grants from the National Key Research and Development Program (2021YFC2300603), the National Natural Science Foundation of China (32000633, 31730026, 81930039), the Natural Science Foundation of Shandong Province (ZR2020QH136), the China Postdoctoral Science Foundation (2020M682187), and the Postdoctoral Innovation Project of Shandong Province (202002012). We thank Dr. Xiaopeng Qi (Shandong University) for the F. novicida strain U112.

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CG conceived the research; CG and FL designed the experiments; WZ and FL performed the research; LZ, YZ, BL, CM, WZ, and SL, provided discussions; CG and FL analyzed the data; CG and FL wrote the paper.

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Correspondence to Feng Liu or Chengjiang Gao.

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The authors declare no competing interests.

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Zhuang, W., Zhang, L., Zheng, Y. et al. USP3 deubiquitinates and stabilizes the adapter protein ASC to regulate inflammasome activation. Cell Mol Immunol 19, 1141–1152 (2022). https://doi.org/10.1038/s41423-022-00917-7

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  • DOI: https://doi.org/10.1038/s41423-022-00917-7

Keywords

  • ASC
  • inflammasome
  • K48-linked ubiquitination
  • proteasomal degradation
  • USP3

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