Abstract

The NLRP3 inflammasome responds to microbes and danger signals by processing and activating proinflammatory cytokines, including interleukin 1β (IL-1β) and IL-18. We found here that activation of the NLRP3 inflammasome was restricted to interphase of the cell cycle by NEK7, a serine-threonine kinase previously linked to mitosis. Activation of the NLRP3 inflammasome required NEK7, which bound to the leucine-rich repeat domain of NLRP3 in a kinase-independent manner downstream of the induction of mitochondrial reactive oxygen species (ROS). This interaction was necessary for the formation of a complex containing NLRP3 and the adaptor ASC, oligomerization of ASC and activation of caspase-1. NEK7 promoted the NLRP3-dependent cellular inflammatory response to intraperitoneal challenge with monosodium urate and the development of experimental autoimmune encephalitis in mice. Our findings suggest that NEK7 serves as a cellular switch that enforces mutual exclusivity of the inflammasome response and cell division.

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References

  1. 1.

    , , & The inflammasome: an integrated view. Immunol. Rev. 243, 136–151 (2011).

  2. 2.

    , , & A role for mitochondria in NLRP3 inflammasome activation. Nature 469, 221–225 (2011).

  3. 3.

    , , , & The adaptor MAVS promotes NLRP3 mitochondrial localization and inflammasome activation. Cell 153, 348–361 (2013).

  4. 4.

    et al. Microtubule-driven spatial arrangement of mitochondria promotes activation of the NLRP3 inflammasome. Nat. Immunol. 14, 454–460 (2013).

  5. 5.

    , , & Nercc1, a mammalian NIMA-family kinase, binds the Ran GTPase and regulates mitotic progression. Genes Dev. 16, 1640–1658 (2002).

  6. 6.

    et al. A mitotic cascade of NIMA family kinases. Nercc1/Nek9 activates the Nek6 and Nek7 kinases. J. Biol. Chem. 278, 34897–34909 (2003).

  7. 7.

    , , & Nek7 kinase is enriched at the centrosome, and is required for proper spindle assembly and mitotic progression. FEBS Lett. 580, 6489–6495 (2006).

  8. 8.

    & The Nek6 and Nek7 protein kinases are required for robust mitotic spindle formation and cytokinesis. Mol. Cell. Biol. 29, 3975–3990 (2009).

  9. 9.

    et al. Nek9 is a Plk1-activated kinase that controls early centrosome separation through Nek6/7 and Eg5. EMBO J. 30, 2634–2647 (2011).

  10. 10.

    , & NEK7 is a centrosomal kinase critical for microtubule nucleation. Biochem. Biophys. Res. Commun. 360, 56–62 (2007).

  11. 11.

    et al. Nek7 kinase targeting leads to early mortality, cytokinesis disturbance and polyploidy. Oncogene 29, 4046–4057 (2010).

  12. 12.

    , & NEK7 is essential for centriole duplication and centrosomal accumulation of pericentriolar material proteins in interphase cells. J. Cell Sci. 124, 3760–3770 (2011).

  13. 13.

    et al. Real-time resolution of point mutations that cause phenovariance in mice. Proc. Natl. Acad. Sci. USA 112, E440–E449 (2015).

  14. 14.

    et al. Non-canonical inflammasome activation targets caspase-11. Nature 479, 117–121 (2011).

  15. 15.

    et al. Experimental autoimmune encephalomyelitis is exacerbated by IL-1α and suppressed by soluble IL-1 receptor. J. Immunol. 146, 2983–2989 (1991).

  16. 16.

    & Protective effect of the interleukin-1 receptor antagonist (IL-1ra) on experimental allergic encephalomyelitis in rats. J. Neuroimmunol. 61, 241–245 (1995).

  17. 17.

    et al. K+ efflux is the common trigger of NLRP3 inflammasome activation by bacterial toxins and particulate matter. Immunity 38, 1142–1153 (2013).

  18. 18.

    et al. The calcium-sensing receptor regulates the NLRP3 inflammasome through Ca2+ and cAMP. Nature 492, 123–127 (2012).

  19. 19.

    et al. The pyroptosome: a supramolecular assembly of ASC dimers mediating inflammatory cell death via caspase-1 activation. Cell Death Differ. 14, 1590–1604 (2007).

  20. 20.

    , , , & Differential requirement for caspase-1 autoproteolysis in pathogen-induced cell death and cytokine processing. Cell Host Microbe 8, 471–483 (2010).

  21. 21.

    et al. Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. Cell 156, 1193–1206 (2014).

  22. 22.

    et al. Cell volume regulation modulates NLRP3 inflammasome activation. Immunity 37, 487–500 (2012).

  23. 23.

    , & P2X7 receptor differentially couples to distinct release pathways for IL-1β in mouse macrophage. J. Immunol. 180, 7147–7157 (2008).

  24. 24.

    , , , & Anakinra therapy for CINCA syndrome with a novel mutation in exon 4 of the CIAS1 gene. Acta Paediatr. 95, 246–249 (2006).

  25. 25.

    , , , & Mutational analysis in neonatal-onset multisystem inflammatory disease: comment on the articles by Frenkel et al. and Saito et al. Arthritis Rheum. 54, 2703–2704 (2006).

  26. 26.

    et al. Phenotype-genotype analysis of cryopyrin-associated periodic syndromes (CAPS): description of a rare non-exon 3 and a novel CIAS1 missense mutation. J. Clin. Immunol. 28, 134–138 (2008).

  27. 27.

    , , , & Isolation and characterization of two evolutionarily conserved murine kinases (Nek6 and Nek7) related to the fungal mitotic regulator, NIMA. Genomics 68, 187–196 (2000).

  28. 28.

    et al. Selective small-molecule inhibitor reveals critical mitotic functions of human CDK1. Proc. Natl. Acad. Sci. USA 103, 10660–10665 (2006).

  29. 29.

    , & Mitochondria in the regulation of innate and adaptive immunity. Immunity 42, 406–417 (2015).

  30. 30.

    in The Cell: A Molecular Approach (Sinauer Associates, Sunderland, Massachusetts, 2000).

  31. 31.

    Mechanisms of interleukin-1β release. Immunobiology 214, 543–553 (2009).

  32. 32.

    & Interleukin-1beta induction of NFκB is partially regulated by H2O2-mediated activation of NFκB-inducing kinase. J. Biol. Chem. 281, 1495–1505 (2006).

  33. 33.

    & Cellular responses to reactive oxygen species-induced DNA damage and aging. Biol. Chem. 389, 211–220 (2008).

  34. 34.

    et al. Mitosis inhibits DNA double-strand break repair to guard against telomere fusions. Science 344, 189–193 (2014).

  35. 35.

    , , & ENU mutagenesis in mice. Methods Mol. Biol. 415, 1–16 (2008).

  36. 36.

    , & Critical role of antigen-specific antibody in experimental autoimmune encephalomyelitis induced by recombinant myelin oligodendrocyte glycoprotein. Eur. J. Immunol. 32, 1905–1913 (2002).

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Acknowledgements

We thank H. Zaki (University of Texas Southwestern Medical Center) for Nlrp3−/− (B6.129S6-Nlrp3tm1Bhk/J) mice; and F. Shao (National Institute of Biological Sciences, Beijing, China) for the NLRC4 plasmid. Supported by the US National Institutes of Health (U19 AI100627).

Author information

Affiliations

  1. Center for the Genetics of Host Defense, University of Texas Southwestern Medical Center, Dallas, Texas, USA.

    • Hexin Shi
    • , Ying Wang
    • , Xiaohong Li
    • , Xiaoming Zhan
    • , Miao Tang
    • , Maggy Fina
    • , Lijing Su
    • , David Pratt
    • , Chun Hui Bu
    • , Sara Hildebrand
    • , Stephen Lyon
    • , Lindsay Scott
    • , Jiexia Quan
    • , Qihua Sun
    • , Jamie Russell
    • , Stephanie Arnett
    • , Peter Jurek
    • , Eva Marie Y Moresco
    •  & Bruce Beutler
  2. Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical Center, Dallas, Texas, USA.

    • Ding Chen
    •  & Nancy L Monson
  3. Department of Immunology and Microbial Sciences, The Scripps Research Institute, La Jolla, California, USA.

    • Vladimir V Kravchenko
    • , John C Mathison
    •  & Richard J Ulevitch

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Contributions

H.S. and B.B. designed the study and analyzed data, with suggestions from N.L.M. and R.J.U.; H.S., Y.W., L.Su, D.C., V.V.K. and J.C.M. performed experiments; H.S. and Y.W. identified the Cuties phenotype; X.L., X.Z., M.T. and M.F. generated the Nek7-knockout mice; D.P., C.H.B., S.H., S.L., L.Sc, J.Q. and Q.S. performed genome mapping and genotyping; H.S., J.R. and S.A. maintained the Cuties mice; H.S., P.J., E.M.Y.M. and B.B. edited the figures; and H.S., E.M.Y.M. and B.B. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Bruce Beutler.

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DOI

https://doi.org/10.1038/ni.3333