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Bcl10 activates the NF-κB pathway through ubiquitination of NEMO

Nature volume 427, pages 167–171 (2004)Cite this article

Abstract

The NF-κB family of transcription factors is activated in response to many stimuli, including pro-inflammatory cytokines, environmental stresses and, in the case of B and T lymphocytes, by antigenic stimulation1,2. Bcl10 is essential for NF-κB activation by T- and B-cell receptors. T and B lymphocytes from Bcl10-deficient mice fail to activate NF-κB in response to antigen-receptor stimulation and, as a consequence, are unable to proliferate3. Bcl10 overexpression is sufficient to activate NF-κB, a process that requires the NF-κB essential modulator NEMO (also known as IKK-γ), which is the regulatory subunit of the IκB kinase complex4. However, the cellular mechanism by which Bcl10 activates the NF-κB pathway remains unclear. Here we show that Bcl10 targets NEMO for lysine-63-linked ubiquitination. Notably, a mutant form of NEMO that cannot be ubiquitinated inhibited Bcl10-induced NF-κB activation. Paracaspase and a ubiquitin-conjugating enzyme (UBC13) were both required for Bcl10-induced NEMO ubiquitination and subsequent NF-κB activation. Furthermore, short interfering RNAs that reduced the expression of paracaspase and UBC13 abrogated the effects of Bcl10. Thus, the adaptor protein Bcl10 promotes activation of NF-κB transcription factors through paracaspase- and UBC13-dependent ubiquitination of NEMO.

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Figure 1: Isolation of a Bcl10 signalling complex and selective requirement of paracaspase for NF-κB activation by Bcl10.
Figure 2: Paracaspase and UBC13 are required for Bcl10-induced ubiquitination and NF-κB activation.
Figure 3: NEMO is a substrate of the Bcl10-induced ubiquitination pathway.
Figure 4: Role of NEMO ubiquitination in NF-κB activation.

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References

  1. Silverman, N. & Maniatis, T. NF-κB signaling pathways in mammalian and insect innate immunity. Genes Dev. 15, 2321–2342 (2001)

    Article  CAS  Google Scholar 

  2. Dixit, V. M. & Mak, T. W. NF-κB signaling: Many roads lead to Madrid. Cell 111, 615–619 (2002)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  4. Lucas, P. C. et al. Bcl10 and MALT1, independent targets of chromosomal translocation in malt lymphoma, cooperate in a novel NF-κB signaling pathway. J. Biol. Chem. 276, 19012–19019 (2001)

    Article  CAS  Google Scholar 

  5. Rothwarf, D. M. & Karin, M. The NF-κB activation pathway: a paradigm in information transfer from membrane to nucleus. Sci. STKE 5, RE1 (1999)

    Google Scholar 

  6. Ben-Neriah, Y. Regulatory functions of ubiquitination in the immune system. Nature Immunol. 3, 20–26 (2002)

    Article  CAS  Google Scholar 

  7. Hoffmann, A., Levchenko, A., Scott, M. L. & Baltimore, D. The IκB-NF-κB signaling module: temporal control and selective gene activation. Science 298, 1241–1245 (2002)

    Article  ADS  CAS  Google Scholar 

  8. Willis, T. G. et al. Bcl10 is involved in t(1;14)(p22;q32) of MALT B cell lymphoma and mutated in multiple tumor types. Cell 96, 35–45 (1999)

    Article  CAS  Google Scholar 

  9. Zhang, Q. et al. Inactivating mutations and overexpression of BCL10, a caspase recruitment domain-containing gene, in MALT lymphoma with t(1;14)(p22;q32). Nature Genet. 22, 63–68 (1999)

    Article  CAS  Google Scholar 

  10. Thome, M. & Tschopp, J. TCR-induced NF-κB activation: a crucial role for Carma1. Bcl10 and MALT1. Trends Immunol. 24, 419–424 (2003)

    Article  CAS  Google Scholar 

  11. Uren, A. G. et al. Identification of paracaspases and metacaspases: two ancient families of caspase-like proteins, one of which plays a key role in MALT lymphoma. Mol. Cell 6, 961–967 (2000)

    CAS  PubMed  Google Scholar 

  12. Shi, C. S. & Kehrl, J. H. Tumor necrosis factor (TNF)-induced germinal center kinase-related (GCKR) and stress-activated protein kinase (SAPK) activation depends upon the E2/E3 complex Ubc13-Uev1A/TNF receptor-associated factor 2 (TRAF2). J. Biol. Chem. 278, 15429–15434 (2003)

    Article  CAS  Google Scholar 

  13. Deng, L. et al. Activation of the IκB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain. Cell 103, 351–361 (2000)

    Article  CAS  Google Scholar 

  14. Weissman, A. M. Themes and variations on ubiquitylation. Nature Rev. Mol. Cell Biol. 2, 169–178 (2001)

    Article  CAS  Google Scholar 

  15. Wang, C. et al. TAK1 is a ubiquitin-dependent kinase of MKK and IKK. Nature 412, 346–351 (2001)

    Article  ADS  CAS  Google Scholar 

  16. Hoege, C. et al. RAD6-dependent DNA repair is linked to modification of PCNA by ubiquitin and SUMO. Nature 419, 135–141 (2002)

    Article  ADS  CAS  Google Scholar 

  17. Makris, C., Roberts, J. L. & Karin, M. The carboxyl-terminal region of IκB kinase gamma (IKKγ) is required for full IKK activation. Mol. Cell. Biol. 22, 6573–6581 (2002)

    Article  CAS  Google Scholar 

  18. Smahi, A. et al. Genomic rearrangement in NEMO impairs NF-B activation and is a cause of incontinentia pigmenti. Nature 405, 466–472 (2000)

    Article  ADS  CAS  Google Scholar 

  19. Yamaoka, S. et al. Complementation cloning of NEMO, a component of the IκB kinase complex essential for NF-κB activation. Cell 93, 1231–1240 (1998)

    Article  CAS  Google Scholar 

  20. Harhaj, E. W. et al. Somatic mutagenesis studies of NF-κB signaling in human T cells: evidence for an essential role of IKKγ in NF-κB activation by T-cell costimulatory signals and HTLV-I Tax protein. Oncogene 19, 1448–1456 (2000)

    Article  CAS  Google Scholar 

  21. Ohi, M. D. et al. Structural insights into the U-box, a domain associated with multi-ubiquitination. Nature Struct. Biol. 10, 250–255 (2003)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  23. Kovalenko, A. et al. The tumour suppressor CYLD negatively regulates NF-B signalling by deubiquitination. Nature 424, 801–805 (2003)

    Article  ADS  CAS  Google Scholar 

  24. Brummelkamp, T. et al. Loss of the cylindromatosis tumour suppressor inhibits apoptosis by activating NF-κB. Nature 424, 797–801 (2003)

    Article  ADS  CAS  Google Scholar 

  25. Trompouki, E. et al. CYLD is a deubiquitinating enzyme that negatively regulates NF-B activation by TNFR family members. Nature 424, 793–796 (2003)

    Article  ADS  CAS  Google Scholar 

  26. Harbury, P. B. et al. A switch between two-, three- and four-stranded coiled coil in GCN4 leucine zipper mutants. Science 262, 1401–1407 (1993)

    Article  ADS  CAS  Google Scholar 

  27. Li, M. et al. Deubiquitination of p53 by HAUSP is an important pathway for p53 stabilization. Nature 416, 648–653 (2002)

    Article  ADS  CAS  Google Scholar 

  28. Moraes, T. F. et al. Crystal structure of the human ubiquitin conjugating enzyme complex, hMms2-hUbc13. Nature Struct. Biol. 8, 669–673 (2001)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank members of the Dixit laboratory for discussions and support; K. Newton for critical reading of the manuscript; S. Sun for the JM4.5.2 cell line; S. Yamaoka for the 5R cell line; R. Baker for Usp2; M. Yan for experimental advice; Z. Zhang for bioinformatics analysis; P. Andersen for antibody preparation; P. Yin for protein purification; and the Genentech Sequencing Core. I.E.W. is funded by a PSTP fellowship from the University of California at Davis.

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Authors and Affiliations

  1. Department of Molecular Oncology, Genentech Inc. 1 DNA Way, South San Francisco, California, 94080, USA

    Honglin Zhou, Ingrid Wertz, Karen O'Rourke & Vishva M. Dixit

  2. Department of Protein Engineering, Genentech Inc. 1 DNA Way, South San Francisco, California, 94080, USA

    Mark Ultsch

  3. Department of Molecular Biology, Genentech Inc. 1 DNA Way, South San Francisco, California, 94080, USA

    Somasekar Seshagiri & Michael Eby

  4. Department of Biological Chemistry, School of Medicine, University of California at Davis, Davis, California, 95616, USA

    Ingrid Wertz

  5. Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, Saskatchewan, S7N 5E5, Canada

    Wei Xiao

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Correspondence to Vishva M. Dixit.

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Supplementary information

Supplementary Figure 1: Isolation of a Bcl10 signalling complex. (JPG 35 kb)

41586_2004_BFnature02273_MOESM2_ESM.jpg

Supplementary Figure 2: a. Paracaspase C-terminus is associated with in vitro ubiquitin ligase activity. In vitro auto-ubiquitination with biotinylated ubiquitin was detected by anti-biotin immunoblotting. b. In vitro auto-ubiquitination of MBP-tagged paracaspase (312-582). Ubiquitination was detected as above. (JPG 44 kb)

41586_2004_BFnature02273_MOESM3_ESM.jpg

Supplementary Figure 3: K399 is conserved in NEMO from multiple species. Shown is an alignment between residues 385 and 419 of human NEMO. K399 is marked by an asterisk. (JPG 84 kb)

Supplementary Figure 4: NEMO K399R mutant does not inhibit NF-kB activation by NIK. (JPG 32 kb)

41586_2004_BFnature02273_MOESM5_ESM.jpg

Supplementary Figure 5: Human paracaspase mutant (C453A) does not inhibit NEMO ubiquitination induced by Bcl10. (JPG 23 kb)

Supplementary Figure Legends (RTF 5 kb)

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Zhou, H., Wertz, I., O'Rourke, K. et al. Bcl10 activates the NF-κB pathway through ubiquitination of NEMO. Nature 427, 167–171 (2004). https://doi.org/10.1038/nature02273

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