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Regulation of Toll/IL-1-receptor-mediated gene expression by the inducible nuclear protein IκBζ


Toll-like receptors (TLRs) recognize microbial components and trigger the inflammatory and immune responses against pathogens. IκBζ (also known as MAIL and INAP) is an ankyrin-repeat-containing nuclear protein that is highly homologous to the IκB family member Bcl-3 (refs 1–6). Transcription of IκBζ is rapidly induced by stimulation with TLR ligands and interleukin-1 (IL-1). Here we show that IκBζ is indispensable for the expression of a subset of genes activated in TLR/IL-1R signalling pathways. IκBζ-deficient cells show severe impairment of IL-6 production in response to a variety of TLR ligands as well as IL-1, but not in response to tumour-necrosis factor-α. Endogenous IκBζ specifically associates with the p50 subunit of NF-κB, and is recruited to the NF-κB binding site of the IL-6 promoter on stimulation. Moreover, NF-κB1/p50-deficient mice show responses to TLR/IL-1R ligands similar to those of IκBζ-deficient mice. Endotoxin-induced expression of other genes such as Il12b and Csf2 is also abrogated in IκBζ-deficient macrophages. Given that the lipopolysaccharide-induced transcription of IκBζ occurs earlier than transcription of these genes, some TLR/IL-1R-mediated responses may be regulated in a gene expression process of at least two steps that requires inducible IκBζ.

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Figure 1: Specific induction of IκBζ on stimulation by TLR/IL-1R ligands.
Figure 2: Immune responses in IκBζ-/- cells and kinetics of IκBζ induction.
Figure 3: In vitro analysis of IκBζ on the Il6 promoter.
Figure 4: The TLR/IL-1R responses in NF-κB1/p50-deficient cells and microarray analysis of IκBζ-/- cells a, NF-κB1+/+ (filled symbols) and NF-κB1-/- (open symbols) peritoneal macrophages were cultured with 10 ng ml-1 LPS in the presence of 30 ng ml-1 IFN-γ for 24 h.
Figure 5: In vivo cytokine production in IκBζ-/- mice.


  1. Takeda, K., Kaisho, T. & Akira, S. Toll-like receptors. Annu. Rev. Immunol. 21, 335–376 (2003)

    CAS  Article  Google Scholar 

  2. Janeway, C. A. Jr & Medzhitov, R. Innate immune recognition. Annu. Rev. Immunol. 20, 197–216 (2002)

    CAS  Article  Google Scholar 

  3. O'Neill, L. A. Therapeutic targeting of Toll-like receptors for inflammatory and infectious diseases. Curr. Opin. Pharmacol. 3, 396–403 (2003)

    CAS  Article  Google Scholar 

  4. Yamazaki, S., Muta, T. & Takeshige, K. A novel IκB protein, IκB-ζ, induced by proinflammatory stimuli, negatively regulates nuclear factor-κB in the nuclei. J. Biol. Chem. 276, 27657–27662 (2001)

    CAS  Article  Google Scholar 

  5. Kitamura, H., Kanehira, K., Okita, K., Morimatsu, M. & Saito, M. MAIL, a novel nuclear IκB protein that potentiates LPS-induced IL-6 production. FEBS Lett. 485, 53–56 (2000)

    CAS  Article  Google Scholar 

  6. Haruta, H., Kato, A. & Todokoro, K. Isolation of a novel interleukin-1-inducible nuclear protein bearing ankyrin-repeat motifs. J. Biol. Chem. 276, 12485–12488 (2001)

    CAS  Article  Google Scholar 

  7. Muzio, M., Ni, J., Feng, P. & Dixit, V. M. IRAK (Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signaling. Science 278, 1612–1615 (1997)

    ADS  CAS  Article  Google Scholar 

  8. Wesche, H., Henzel, W. J., Shillinglaw, W., Li, S. & Cao, Z. MyD88: an adapter that recruits IRAK to the IL-1 receptor complex. Immunity 7, 837–847 (1997)

    CAS  Article  Google Scholar 

  9. Medzhitov, R. et al. MyD88 is an adaptor protein in the hToll/IL-1 receptor family signaling pathways. Mol. Cell 2, 253–258 (1998)

    CAS  Article  Google Scholar 

  10. Alexopoulou, L., Holt, A. C., Medzhitov, R. & Flavell, R. A. Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3. Nature 413, 732–738 (2001)

    ADS  CAS  Article  Google Scholar 

  11. Kawai, T. et al. Lipopolysaccharide stimulates the MyD88-independent pathway and results in activation of IFN-regulatory factor 3 and the expression of a subset of lipopolysaccharide-inducible genes. J. Immunol. 167, 5887–5894 (2001)

    CAS  Article  Google Scholar 

  12. Doyle, S. et al. IRF3 mediates a TLR3/TLR4-specific antiviral gene program. Immunity 17, 251–263 (2002)

    CAS  Article  Google Scholar 

  13. Ghosh, S. & Karin, M. Missing pieces in the NF-κB puzzle. Cell 109(suppl.), S81–S96 (2002)

    CAS  Article  Google Scholar 

  14. Zhang, G. & Ghosh, S. Molecular mechanisms of NF-κB activation induced by bacterial lipopolysaccharide through Toll-like receptors. J. Endotoxin Res. 6, 453–457 (2000)

    CAS  Article  Google Scholar 

  15. Janssens, S. & Beyaert, R. Functional diversity and regulation of different interleukin-1 receptor-associated kinase (IRAK) family members. Mol. Cell 11, 293–302 (2003)

    CAS  Article  Google Scholar 

  16. Eto, A., Muta, T., Yamazaki, S. & Takeshige, K. Essential roles for NF-κB and a Toll/IL-1 receptor domain-specific signal(s) in the induction of IκB-ζ. Biochem. Biophys. Res. Commun. 301, 495–501 (2003)

    CAS  Article  Google Scholar 

  17. Akira, S., Taga, T. & Kishimoto, T. Interleukin-6 in biology and medicine. Adv. Immunol. 54, 1–78 (1993)

    CAS  Article  Google Scholar 

  18. Matsusaka, T. et al. Transcription factors NF-IL6 and NF-κB synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. Proc. Natl Acad. Sci. USA 90, 10193–10197 (1993)

    ADS  CAS  Article  Google Scholar 

  19. Zhong, H., May, M. J., Jimi, E. & Ghosh, S. The phosphorylation status of nuclear NF-κB determines its association with CBP/p300 or HDAC-1. Mol. Cell 9, 625–636 (2002)

    CAS  Article  Google Scholar 

  20. Sha, W. C., Liou, H. C., Tuomanen, E. I. & Baltimore, D. Targeted disruption of the p50 subunit of NF-κB leads to multifocal defects in immune responses. Cell 80, 321–330 (1995)

    CAS  Article  Google Scholar 

  21. Sanjabi, S., Hoffmann, A., Liou, H. C., Baltimore, D. & Smale, S. T. Selective requirement for c-Rel during IL-12 P40 gene induction in macrophages. Proc. Natl Acad. Sci. USA 97, 12705–12710 (2000)

    ADS  CAS  Article  Google Scholar 

  22. Hoffmann, A., Leung, T. H. & Baltimore, D. Genetic analysis of NF-κB/Rel transcription factors defines functional specificities. EMBO J. 22, 5530–5539 (2003)

    CAS  Article  Google Scholar 

  23. Hill, M. R. & McCallum, R. E. Identification of tumor necrosis factor as a transcriptional regulator of the phosphoenolpyruvate carboxykinase gene following endotoxin treatment of mice. Infect. Immun. 60, 4040–4050 (1992)

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Pasparakis, M. et al. TNF-mediated inflammatory skin disease in mice with epidermis-specific deletion of IKK2. Nature 417, 861–866 (2002)

    ADS  CAS  Article  Google Scholar 

  25. Douni, E. et al. Transgenic and knockout analyses of the role of TNF in immune regulation and disease pathogenesis. J. Inflamm. 47, 27–38 (1995)

    CAS  PubMed  Google Scholar 

  26. Yamamoto, M. et al. Role of adaptor TRIF in the MyD88-independent Toll-Like Receptor signaling pathway. Science 301, 640–643 (2003)

    ADS  CAS  Article  Google Scholar 

  27. Kinoshita, S., Akira, S. & Kishimoto, T. A member of the C/EBP family, NF-IL6 beta, forms a heterodimer and transcriptionally synergizes with NF-IL6. Proc. Natl Acad. Sci. USA 89, 1473–1476 (1992)

    ADS  CAS  Article  Google Scholar 

  28. Saitoh, T. et al. TWEAK induces NF-κB2 p100 processing and long lasting NF-κB activation. J. Biol. Chem. 278, 36005–36012 (2003)

    CAS  Article  Google Scholar 

  29. Toshchakov, V. et al. TLR4, but not TLR2, mediates IFN-β-induced STAT1α/β-dependent gene expression in macrophages. Nature Immunol. 3, 392–398 (2002)

    CAS  Article  Google Scholar 

  30. Kuwata, H. et al. IL-10-inducible Bcl-3 negatively regulates LPS-induced TNF-α production in macrophages. Blood 102, 4123–4129 (2003)

    CAS  Article  Google Scholar 

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We thank T. Kitamura, A. Aderem, D. Golenbock and H. Tomizawa for providing Plat-E packaging cell lines, flagellin, the ELAM1 reporter plasmid, and R-848, respectively. We also thank T. Kawai and K. Ishii for discussions; M. Hashimoto for secretarial assistance; and N. Okita and N. Iwami for technical assistance. This work was supported by grants from Special Coordination Funds, the Ministry of Education, Culture, Sports, Science and Technology, Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists, The Uehara Memorial Foundation, The Naito Foundation, and The Junior Research Associate from RIKEN.

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Correspondence to Shizuo Akira.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Discussion 1

Targeted Disruption of murine IκB-ζ gene. (DOC 20 kb)

Supplementary Discussion 2

Relation between the κB sequences of the gene tested and IκB-ζ requirement and regulation of IκB-ζ expression. (DOC 20 kb)

Supplementary Figure 1

Targeted Disruption of murine IκB-ζ gene. (JPG 82 kb)

Supplementary Figure 2

Histological analysis on IκB-ζ-/- mice. (JPG 164 kb)

Supplementary Figure 3

IL-1β-induced activation of signaling cascades IκB-ζ-/- MEFs. (JPG 50 kb)

Supplementary Figure 4a

Gene expression analysis of LPS-inducible genes with statistically selected genes. (JPG 154 kb)

Supplementary Figure 4b-d

Gene expression analysis of LPS-inducible genes with statistically selected genes. (JPG 89 kb)

Supplementary Figure Legends

(DOC 27 kb)

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Yamamoto, M., Yamazaki, S., Uematsu, S. et al. Regulation of Toll/IL-1-receptor-mediated gene expression by the inducible nuclear protein IκBζ. Nature 430, 218–222 (2004).

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