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Involvement of linear polyubiquitylation of NEMO in NF-κB activation

Nature Cell Biology volume 11, pages 123–132 (2009)Cite this article

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Abstract

Nuclear factor-κB (NF-κB) is a key transcription factor in inflammatory, anti-apoptotic and immune processes. The ubiquitin pathway is crucial in regulating the NF-κB pathway. We have found that the LUBAC ligase complex, composed of the two RING finger proteins HOIL-1L and HOIP, conjugates a head-to-tail-linked linear polyubiquitin chain to substrates. Here, we demonstrate that LUBAC activates the canonical NF-κB pathway by binding to NEMO (NF-κB essential modulator, also called IKKγ) and conjugates linear polyubiquitin chains onto specific Lys residues in the CC2–LZ domain of NEMO in a Ubc13-independent manner. Moreover, in HOIL-1 knockout mice and cells derived from these mice, NF-κB signalling induced by pro-inflammatory cytokines such as TNF-α and IL-1β was suppressed, resulting in enhanced TNF-α–induced apoptosis in hepatocytes of HOIL-1 knockout mice. These results indicate that LUBAC is involved in the physiological regulation of the canonical NF-κB activation pathway through linear polyubiquitylation of NEMO.

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Figure 1: LUBAC activates the NF-κB pathway.
Figure 2: Linear polyubiquitylation activity of LUBAC is indispensable for canonical NF-κB activation.
Figure 3: LUBAC targets NEMO for NF-κB activation.
Figure 4: LUBAC conjugates a linear polyubiquitin chain to NEMO.
Figure 5: Lys 285 and Lys 309 in the CC2∼LZ domain of NEMO are acceptor sites for LUBAC-mediated linear polyubiquitylation.
Figure 6: Ubc13-independent NF-κB activation by LUBAC.
Figure 7: Impaired TNF-α signalling in HOIL-1 null cells and mice.

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Change history

  • 13 January 2009

    In the version of this article initially published online figure panels 3a, 3e and 7e were not clearly defined. Also (NEMOCC2~LZ) was corrected to (NEMO∆CC2~LZ). These errors have been corrected for the print, HTML and PDF versions of the article.

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Acknowledgements

We would like to thank Nobuhito Goda and Natsuko Fujiki for instruction in mouse primary hepatocyte preparation; Ayumi Taya for supporting the LC and MS analyses; and Koichi Nakajima for reagents. This work was partly supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan (F.T., T.T. and K.I.), the Japan Health Sciences Foundation (F.T.) and the Naito Foundation (F.T.).

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Authors and Affiliations

  1. Department of Biophysics and Biochemistry, Graduate School of Medicine and Cell Biology and Metabolism Group, Graduate School of Frontier Biosciences, Osaka University, Suita, 565-0871, Osaka, Japan

    Fuminori Tokunaga, Shin-ichi Sakata, Kiyoko Kamei, Tomoko Nakagawa & Kazuhiro Iwai

  2. CREST, Japan Science Technology Corporation, Kawaguchi, 332-0012, Saitama, Japan

    Fuminori Tokunaga, Shin-ichi Sakata & Kazuhiro Iwai

  3. Department of Molecular Cell Biology, Graduate School of Medicine, Osaka City University, Abeno-ku, 545-8585, Osaka

    Fuminori Tokunaga, Shin-ichi Sakata, Takayoshi Kirisako, Kiyoko Kamei, Tomoko Nakagawa, Michiko Kato & Kazuhiro Iwai

  4. Laboratory of Frontier Science, Core Technology and Research Center, Tokyo Metropolitan Institute of Medical Science, Bunkyo-ku, 113-8613, Tokyo, Japan

    Yasushi Saeki, Shigeo Murata & Keiji Tanaka

  5. Laboratory of Protein Profiling and Functional Proteomics, Institute for Protein Research, Osaka University, Suita, 565-0871, Osaka, Japan

    Yoshinori Satomi & Toshifumi Takao

  6. Laboratory of Protein Metabolism, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Bunkyo-ku, 113-0033, Tokyo, Japan

    Shigeo Murata

  7. Department of Molecular Virology, Graduate School of Medicine, Tokyo Medical and Dental University, Bunkyo-ku, 113-8519, Tokyo, Japan

    Shoji Yamaoka

  8. Department of Microbiology and Immunology, Laboratory of Immune Regulation, Graduate School of Medicine,

    Masahiro Yamamoto

  9. Laboratory of Host Defense, WPI Immunology Frontier Research Center, Osaka University, Suita, 565-0871, Osaka, Japan

    Shizuo Akira

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  1. Fuminori Tokunaga
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Contributions

F.T., S.-I.S, K.K., T.N. and M.K. performed experiments; Y.S., Y.S. and T.T. conducted mass spectrometric analyses. S.-I.S. and S.M. generated the HOIL-1 KO mice; T.K., S.Y., M.Y., S.A. and K.T. provided reagents and suggestions; F.T. and K.I. coordinated the study and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Kazuhiro Iwai.

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Tokunaga, F., Sakata, Si., Saeki, Y. et al. Involvement of linear polyubiquitylation of NEMO in NF-κB activation. Nat Cell Biol 11, 123–132 (2009). https://doi.org/10.1038/ncb1821

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