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Proteolysis of NF-κB1 p105 is essential for T cell antigen receptor–induced proliferation

Nature Immunology volume 10, pages 3847 (2009) | Download Citation

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Abstract

To investigate the importance of proteolysis of NF-κB1 p105 induced by the kinase IKK in activation of the transcription factor NF-κB, we generated 'Nfkb1SSAA/SSAA' mice, in which the IKK-target serine residues of p105 were substituted with alanine. Nfkb1SSAA/SSAA mice had far fewer CD4+ regulatory and memory T cells because of cell-autonomous defects. These T cell subtypes require activation of NF-κB by the T cell antigen receptor for their generation, and the Nfkb1SSAA mutation resulted in less activation of NF-κB in CD4+ T cells and proliferation of CD4+ T cells after stimulation of the T cell antigen receptor. The Nfkb1SSAA mutation also blocked the ability of CD4+ T cells to provide help to wild-type B cells during a primary antibody response. IKK-induced p105 proteolysis is therefore essential for optimal T cell antigen receptor–induced activation of NF-κB and mature CD4+ T cell function.

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References

  1. 1.

    & NF-κB regulation in the immune system. Nat. Rev. Immunol. 2, 725–734 (2002).

  2. 2.

    , & Structure, regulation and function of NF-κB. Annu. Rev. Cell Biol. 10, 405–455 (1994).

  3. 3.

    & Functions of NF-κB1 and NF-κB2 in immune cell biology. Biochem. J. 382, 393–409 (2004).

  4. 4.

    & Phosphorylation meets ubiquitination: the control of NF-κB activity. Annu. Rev. Immunol. 18, 621–663 (2000).

  5. 5.

    et al. Structural motifs involves in ubiquitin-mediated processing of the NF-κB precursor p105: roles of the glycine-rich region and a downstream ubiquitination domain. Mol. Cell. Biol. 19, 3664–3673 (1999).

  6. 6.

    βTrCP-mediated proteolysis of NF-κB1 p105 requires phosphorylation of p105 serines 927 and 932. Mol. Cell. Biol. 23, 402–413 (2003).

  7. 7.

    , , & Shared pathways of Iκβ kinase-induced SCFβTrCP-mediated ubiquitination and degradation for the NF-κB precursor p105 and IκBα. Mol. Cell. Biol. 21, 1024–1035 (2001).

  8. 8.

    et al. Direct phosphorylation of NF-κB p105 by the IκB kinase complex on serine 927 is essential for signal-induced p105 proteolysis. J. Biol. Chem. 276, 22215–22222 (2001).

  9. 9.

    et al. Lipopolysaccharide activation of the TPL-2/MEK/extracellular signal-regulated kinase mitogen-activated protein kinase cascade is regulated by IκB kinase-induced proteolysis of NF-κB1 p105. Mol. Cell. Biol. 24, 9658–9667 (2004).

  10. 10.

    , , , & IKKβ is an essential component of the TPL-2 signaling pathway. Mol. Cell. Biol. 24, 6040–6048 (2004).

  11. 11.

    , & MAP kinase kinase kinases and innate immunity. Trends Immunol. 27, 40–48 (2006).

  12. 12.

    et al. Differential dependence of CD4+ CD25+ regulatory and natural killer-like T cells on signals leading to NF-κB activation. Proc. Natl. Acad. Sci. USA 101, 4566–4571 (2004).

  13. 13.

    et al. Mature T cells depend on signaling through the IKK complex. Immunity 19, 377–389 (2003).

  14. 14.

    et al. Coordination between NF-κB family members p50 and p52 is essential for mediating LTβR signals in the development and organization of secondary lymphoid tissues. Blood 107, 1048–1055 (2006).

  15. 15.

    & A well adapted reguatory contrivance: regulatory T cell development and the forkhead family of transcription factor Foxp3. Nat. Immunol. 6, 331–337 (2005).

  16. 16.

    , , , & In vivo identification of glycolipid antigen-specific T cells using fluorescent CD1d tetramers. J. Exp. Med. 191, 1895–1903 (2000).

  17. 17.

    & Similarities and differences in CD4+ and CD8+ effector and memory T cell generation. Nat. Immunol. 4, 835–842 (2004).

  18. 18.

    , , & Interleukin-2 receptor signaling in regulatory T cell development and homeostasis. Immunol. Lett. 114, 1–8 (2007).

  19. 19.

    et al. In vivo regulation of interleukin-2 receptor α gene transcription by the coordinated binding of constitutive and inducible factors in human primary T cells. EMBO J. 14, 5060–5072 (1995).

  20. 20.

    et al. TNFα induction by LPS is regulated post-transcriptionally via a TPL2/ERK-dependent pathway. Cell 103, 1071–1083 (2000).

  21. 21.

    et al. ABIN-2 forms a ternary complex with TPL-2 and NF-κB1 p105 and is essential for TPL-2 protein stability. Mol. Cell. Biol. 24, 5235–5248 (2004).

  22. 22.

    et al. Chronic inflammation and susceptibility to bacterial infections in mice lacking the polypeptide (p) 105 precursor (NF-κB1) but expressing p50. J. Exp. Med. 187, 985–996 (1998).

  23. 23.

    , , & p105 and p98 precursor proteins play an active role in NF-κB-mediated signal transduction. Genes Dev. 7, 705–718 (1993).

  24. 24.

    , & The precursor of NF-κB p50 has IκB-like functions. Cell 71, 243–253 (1992).

  25. 25.

    , , , & The NF-κB p50 precursor, p105, contains an internal IκB-like inhibitor that preferentially inhibits p50. EMBO J. 11, 3003–3009 (1992).

  26. 26.

    & p105 IκBγ and prototypical IκBs use a similar mechanism to bind but a different mechanism to regulate the subcellular localization of NF-κB. J. Biol. Chem. 278, 556–566 (2003).

  27. 27.

    , & The NF-κB precursor p105 and the proto-oncogene product Bcl-3 are IκB molecules and control nuclear translocation of NF-κB. EMBO J. 12, 213–222 (1993).

  28. 28.

    et al. A fourth IκB protein within the NF-κB signaling module. Cell 128, 369–381 (2007).

  29. 29.

    , , , & Severe liver degeneration in mice lacking the IκB kinase 2 gene. Science 284, 321–325 (1999).

  30. 30.

    et al. Severe liver degeneration and lack of NF-κB activation in NEMO/IKKγ-deficient mice. Genes Dev. 14, 854–862 (2000).

  31. 31.

    et al. Activation by IKKα of a second evolutionary conserved, NF-κB signaling pathway. Science 293, 1495–1499 (2001).

  32. 32.

    et al. BAFF/BLys receptor 3 binds the B cell survival factor BAFF ligand through a discrete surface loop and promotes processing of NF-κB2. Immunity 17, 515–524 (2002).

  33. 33.

    , & Peripheral T cell survival requires continual ligation of the T cell receptor to major histocompatibility complex-encoded molecules. J. Exp. Med. 186, 1269–1275 (1997).

  34. 34.

    et al. Mice lacking the c-Rel proto-oncogene exhibit defects in lymphocyte proliferation, humoral immunity and interleukin-2 expression. Genes Dev. 9, 1965–1977 (1995).

  35. 35.

    , , & NF-κB/Rel participation in the lymphokine-dependent proliferation of T lymphoid cells. J. Immunol. 166, 2218–2227 (2001).

  36. 36.

    et al. IκB kinase 2/β deficiency controls expansion of autoreactive T cells and suppress experimental autoimmune encephalomyelitis. J. Immunol. 179, 179–185 (2007).

  37. 37.

    et al. IκB kinase 2 deficiency in T cells leads to defects in priming, B cell help, germinal center reactions, and homeostatic expansion. J. Immunol. 173, 1612–1619 (2004).

  38. 38.

    et al. Bcl10 is a positive regulator of antigen receptor-induced activation of NF-κB and neural tube closure. Cell 104, 33–42 (2001).

  39. 39.

    , , , & Combined deficiency of p50 and cRel in CD4+ T cells reveals an essential requirement for nuclear factor κB in regulating mature T cell survival and in vivo function. J. Exp. Med. 197, 861–874 (2003).

  40. 40.

    , , , & IL-2 receptor β-dependent STAT5 activation is required for the development of Foxp3+ regulatory cells. J. Immunol. 178, 280–290 (2007).

  41. 41.

    , , & CD28 costimulation of developing thymocytes induces Foxp3 expression and regulatory T cell differentiation independently of interleukin 2. Nat. Immunol. 6, 152–162 (2005).

  42. 42.

    , , & Interleukin-2 enhances CD4+ T cell memory by promoting the generation of IL-7Rα-expressing cells. J. Exp. Med. 204, 547–557 (2007).

  43. 43.

    et al. ABIN-2 is required for optimal activation of the TPL-2/Erk MAP kinase pathway in innate immune responses. Nat. Immunol. 7, 606–615 (2006).

  44. 44.

    , , & NF-κB p105 is a target on IκB kinases and controls signal induction of BCL-3-p50 complexes. EMBO J. 18, 4766–4788 (1999).

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Acknowledgements

We thank P. Tsichlis (Tufts University) and Thomas Jefferson University for Map3k8−/− mice; B. Stockinger and A. O'Garra for advice and critical reading of the manuscript; H. Allen for encouragement at the start of the project; G. Kassiotis for advice on assays for T cell helper function; and the National Institute for Medical Research PhotoGraphics section, Biological Services and flow cytometry service and members of the Ley laboratory for help during the course of this work. Supported by the UK Medical Research Council.

Author information

Author notes

    • Monica P Belich

    Present address: GSK Medicines Research Centre, Stevenage SG1 2NY, UK.

    • Srividya Sriskantharajah
    •  & Monica P Belich

    These authors contributed equally to this work.

Affiliations

  1. Division of Immune Cell Biology, National Institute for Medical Research, London NW7 1AA, UK.

    • Srividya Sriskantharajah
    • , Monica P Belich
    • , Stamatia Papoutsopoulou
    • , Julia Janzen
    • , Victor Tybulewicz
    • , Benedict Seddon
    •  & Steven C Ley

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Contributions

S.S. did experiments for Figures 1, 2, 3, 4, 5, 6, 7, 8; M.P.B. generated the Nfkb1SSAA mutant mouse strain and did experiments for Figure 9; S.P. did experiments for Figures 4e,f and 6e,f; J.J. assayed mRNA by quantitative RT-PCR and IL-2 by ELISA and provided technical support for many experiments; V.T. advised in the design of the Nfkb1SSAA targeting construct and the generation of mutant mouse strains; B.S. helped with the experimental design and several in vivo experiments; and S.C.L. designed the experiments in conjunction with S.S., M.P.B., S.P., J.J. and B.S. and wrote the manuscript.

Corresponding author

Correspondence to Steven C Ley.

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DOI

https://doi.org/10.1038/ni.1685

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