nature.com

SHARPIN is a component of the NF-κB-activating linear ubiquitin chain assembly complex - Nature

  • ️Iwai, Kazuhiro
  • ️Wed Mar 30 2011
  • Letter
  • Published: 30 March 2011

Nature volume 471, pages 633–636 (2011)Cite this article

Subjects

Abstract

Cpdm (chronic proliferative dermatitis) mice develop chronic dermatitis and an immunodeficiency with increased serum IgM1,2,3, symptoms that resemble those of patients with X-linked hyper-IgM syndrome and hypohydrotic ectodermal dysplasia (XHM-ED), which is caused by mutations in NEMO (NF-κB essential modulator; also known as IKBKG)4,5,6. Spontaneous null mutations in the Sharpin (SHANK-associated RH domain interacting protein in postsynaptic density)7 gene are responsible for the cpdm phenotype in mice8. SHARPIN shows significant similarity to HOIL-1L (also known as RBCK1)8,9, a component of linear ubiquitin chain assembly complex (LUBAC), which induces NF-κB activation through conjugation of linear polyubiquitin chains to NEMO10,11,12,13. Here, we identify SHARPIN as an additional component of LUBAC. SHARPIN-containing complexes can linearly ubiquitinate NEMO and activated NF-κB. Thus, we re-define LUBAC as a complex containing SHARPIN, HOIL-1L, and HOIP (also known as RNF31). Deletion of SHARPIN drastically reduced the amount of LUBAC, which resulted in attenuated TNF-α- and CD40-mediated activation of NF-κB in mouse embryonic fibroblasts (MEFs) or B cells from cpdm mice. Considering the pleomorphic phenotype of cpdm mice, these results confirm the predicted role of LUBAC-mediated linear polyubiquitination in NF-κB activation induced by various stimuli, and strongly suggest the involvement of LUBAC-induced NF-κB activation in various disorders.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

References

  1. Gallardo Torres, H. I., Gijbels, M. J., HogenEsch, H. & Kraal, G. Chronic proliferative dermatitis in mice: neutrophil-endothelium interactions and the role of adhesion molecules. Pathobiology 63, 341–347 (1995)

    Article  CAS  Google Scholar 

  2. HogenEsch, H., Janke, S., Boggess, D. & Sundberg, J. P. Absence of Peyer’s patches and abnormal lymphoid architecture in chronic proliferative dermatitis (cpdm/cpdm) mice. J. Immunol. 162, 3890–3896 (1999)

    CAS  PubMed  Google Scholar 

  3. Seymour, R., Sundberg, J. P. & HogenEsch, H. Abnormal lymphoid organ development in immunodeficient mutant mice. Vet. Pathol. 43, 401–423 (2006)

    Article  CAS  Google Scholar 

  4. Zonana, J. et al. A novel X-linked disorder of immune deficiency and hypohidrotic ectodermal dysplasia is allelic to incontinentia pigmenti and due to mutations in IKK-gamma (NEMO). Am. J. Hum. Genet. 67, 1555–1562 (2000)

    Article  CAS  Google Scholar 

  5. Jain, A. et al. Specific missense mutations in NEMO result in hyper-IgM syndrome with hypohydrotic ectodermal dysplasia. Nature Immunol. 2, 223–228 (2001)

    Article  CAS  Google Scholar 

  6. Jain, A. et al. Specific NEMO mutations impair CD40-mediated c-Rel activation and B cell terminal differentiation. J. Clin. Invest. 114, 1593–1602 (2004)

    Article  CAS  Google Scholar 

  7. Lim, S. et al. Sharpin, a novel postsynaptic density protein that directly interacts with the shank family of proteins. Mol. Cell. Neurosci. 17, 385–397 (2001)

    Article  CAS  Google Scholar 

  8. Seymour, R. E. et al. Spontaneous mutations in the mouse Sharpin gene result in multiorgan inflammation, immune system dysregulation and dermatitis. Genes Immun. 8, 416–421 (2007)

    Article  CAS  Google Scholar 

  9. Yamanaka, K. et al. Identification of the ubiquitin-protein ligase that recognizes oxidized IRP2. Nature Cell Biol. 5, 336–340 (2003)

    Article  CAS  Google Scholar 

  10. Kirisako, T. et al. A ubiquitin ligase complex assembles linear polyubiquitin chains. EMBO J. 25, 4877–4887 (2006)

    Article  CAS  Google Scholar 

  11. Tokunaga, F. et al. Involvement of linear polyubiquitylation of NEMO in NF-κB activation. Nature Cell Biol. 11, 123–132 (2009)

    Article  CAS  Google Scholar 

  12. Iwai, K. & Tokunaga, F. Linear polyubiquitination: a new regulator of NF-κB activation. EMBO Rep. 10, 706–713 (2009)

    Article  CAS  Google Scholar 

  13. Haas, T. L. et al. Recruitment of the linear ubiquitin chain assembly complex stabilizes the TNF-R1 signaling complex and is required for TNF-mediated gene induction. Mol. Cell 36, 831–844 (2009)

    Article  CAS  Google Scholar 

  14. Karin, M. Nuclear factor-κB in cancer development and progression. Nature 441, 431–436 (2006)

    Article  ADS  CAS  Google Scholar 

  15. Vallabhapurapu, S. & Karin, M. Regulation and function of NF-κB transcription factors in the immune system. Annu. Rev. Immunol. 27, 693–733 (2009)

    Article  CAS  Google Scholar 

  16. Bhoj, V. G. & Chen, Z. J. Ubiquitylation in innate and adaptive immunity. Nature 458, 430–437 (2009)

    Article  ADS  CAS  Google Scholar 

  17. 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 

  18. Hayden, M. S. & Ghosh, S. Shared principles in NF-κB signaling. Cell 132, 344–362 (2008)

    Article  CAS  Google Scholar 

  19. Gerondakis, S. et al. Unravelling the complexities of the NF-κB signalling pathway using mouse knockout and transgenic models. Oncogene 25, 6781–6799 (2006)

    Article  CAS  Google Scholar 

  20. Piao, J. H. et al. TNF receptor-associated factor 2-dependent canonical pathway is crucial for the development of Peyer’s patches. J. Immunol. 178, 2272–2277 (2007)

    Article  CAS  Google Scholar 

  21. Koni, P. A. et al. Distinct roles in lymphoid organogenesis for lymphotoxins α and β revealed in lymphotoxin β-deficient mice. Immunity 6, 491–500 (1997)

    Article  CAS  Google Scholar 

  22. Xu, M., Skaug, B., Zeng, W. & Chen, Z. J. A Ubiquitin replacement strategy in human cells reveals distinct mechanisms of IKK activation by TNFα and IL-1β. Mol. Cell 36, 302–314 (2009)

    Article  CAS  Google Scholar 

  23. Renninger, M. L., Seymour, R. E., Whiteley, L. O., Sundberg, J. P. & HogenEsch, H. Anti-IL5 decreases the number of eosinophils but not the severity of dermatitis in Sharpin-deficient mice. Exp. Dermatol. 19, 252–258 (2009)

    Article  Google Scholar 

  24. Smahi, A. et al. The NF-κB signalling pathway in human diseases: from incontinentia pigmenti to ectodermal dysplasias and immune-deficiency syndromes. Hum. Mol. Genet. 11, 2371–2375 (2002)

    Article  CAS  Google Scholar 

  25. Jung, J. et al. Newly identified tumor-associated role of human Sharpin. Mol. Cell. Biochem. 340, 161–167 (2010)

    Article  CAS  Google Scholar 

  26. Iwai, K. et al. Identification of the von Hippel–Lindau tumor-suppressor protein as part of an active E3 ubiquitin ligase complex. Proc. Natl Acad. Sci. USA 96, 12436–12441 (1999)

    Article  ADS  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Kitamura, S. Sugano, P. Rennert and J. Browning for reagents; O. Takeuchi for comments; and K. Kamei and K. Fukunaga for technical assistance. This work was partly supported by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (F.T. and K.I.).

Author information

Author notes

  1. Fuminori Tokunaga and Tomoko Nakagawa: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biophysics and Biochemistry, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan,

    Fuminori Tokunaga, Tomoko Nakagawa, Masaki Nakahara, Masami Taniguchi, Shin-ichi Sakata & Kazuhiro Iwai

  2. Laboratory of Protein Metabolism, Tokyo Metropolitan Institute of Medical Science, Setagaya-ku, Tokyo 156-8506, Japan,

    Yasushi Saeki & Keiji Tanaka

  3. Department of Immunology, Juntendo University School of Medicine, Bunkyo-ku, Tokyo 113-8421, Japan,

    Hiroyasu Nakano

  4. Cell Biology and Metabolism Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan,

    Kazuhiro Iwai

Authors

  1. Fuminori Tokunaga

    You can also search for this author in PubMed Google Scholar

  2. Tomoko Nakagawa

    You can also search for this author in PubMed Google Scholar

  3. Masaki Nakahara

    You can also search for this author in PubMed Google Scholar

  4. Yasushi Saeki

    You can also search for this author in PubMed Google Scholar

  5. Masami Taniguchi

    You can also search for this author in PubMed Google Scholar

  6. Shin-ichi Sakata

    You can also search for this author in PubMed Google Scholar

  7. Keiji Tanaka

    You can also search for this author in PubMed Google Scholar

  8. Hiroyasu Nakano

    You can also search for this author in PubMed Google Scholar

  9. Kazuhiro Iwai

    You can also search for this author in PubMed Google Scholar

Contributions

F.T., T.N., M.N., Y.S, M.T., S.-i.S. and H.N. performed experiments. K.T. and K.I. coordinated the study, and F.T. and K.I. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Kazuhiro Iwai.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

PowerPoint slides

About this article

Cite this article

Tokunaga, F., Nakagawa, T., Nakahara, M. et al. SHARPIN is a component of the NF-κB-activating linear ubiquitin chain assembly complex. Nature 471, 633–636 (2011). https://doi.org/10.1038/nature09815

Download citation

  • Received: 25 January 2010

  • Accepted: 11 January 2011

  • Published: 30 March 2011

  • Issue Date: 31 March 2011

  • DOI: https://doi.org/10.1038/nature09815

Editorial Summary

SHARPIN protein role in immune signalling

The ubiquitin conjugation system regulates the canonical (or classical) nuclear factor κB (NF-κB)-activation pathway that mediates immune responses. Linear polyubiquitin chains, in which the C-terminal glycine of ubiquitin is conjugated to the amino group of the N-terminal methionine of another ubiquitin, is generated by a unique ubiquitin ligase complex called LUBAC — the linear ubiquitin chain assembly complex. LUBAC is composed of two RING domain proteins called HOIL-1 and HOIP. Now, three complementary studies published by the laboratories of Henning Walczak, Kazuhiro Iwai and Ivan Dikic identify a novel component of the LUBAC complex called SHARPIN, which is recruited to receptor signalling complexes that form following TNF and CD40L stimulation. The LUBAC complex containing SHARPIN stimulates the formation of linear ubiquitin chains in vitro and in vivo and is required for the activation of NF-κB signalling.