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
Similar content being viewed by others
References
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)
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)
Seymour, R., Sundberg, J. P. & HogenEsch, H. Abnormal lymphoid organ development in immunodeficient mutant mice. Vet. Pathol. 43, 401–423 (2006)
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)
Jain, A. et al. Specific missense mutations in NEMO result in hyper-IgM syndrome with hypohydrotic ectodermal dysplasia. Nature Immunol. 2, 223–228 (2001)
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)
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)
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)
Yamanaka, K. et al. Identification of the ubiquitin-protein ligase that recognizes oxidized IRP2. Nature Cell Biol. 5, 336–340 (2003)
Kirisako, T. et al. A ubiquitin ligase complex assembles linear polyubiquitin chains. EMBO J. 25, 4877–4887 (2006)
Tokunaga, F. et al. Involvement of linear polyubiquitylation of NEMO in NF-κB activation. Nature Cell Biol. 11, 123–132 (2009)
Iwai, K. & Tokunaga, F. Linear polyubiquitination: a new regulator of NF-κB activation. EMBO Rep. 10, 706–713 (2009)
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)
Karin, M. Nuclear factor-κB in cancer development and progression. Nature 441, 431–436 (2006)
Vallabhapurapu, S. & Karin, M. Regulation and function of NF-κB transcription factors in the immune system. Annu. Rev. Immunol. 27, 693–733 (2009)
Bhoj, V. G. & Chen, Z. J. Ubiquitylation in innate and adaptive immunity. Nature 458, 430–437 (2009)
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)
Hayden, M. S. & Ghosh, S. Shared principles in NF-κB signaling. Cell 132, 344–362 (2008)
Gerondakis, S. et al. Unravelling the complexities of the NF-κB signalling pathway using mouse knockout and transgenic models. Oncogene 25, 6781–6799 (2006)
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)
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)
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)
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)
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)
Jung, J. et al. Newly identified tumor-associated role of human Sharpin. Mol. Cell. Biochem. 340, 161–167 (2010)
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)
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
Authors and Affiliations
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
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Figures
This file contains Supplementary Figures 1-22 with legends. (PDF 7646 kb)
Rights and permissions
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature09815
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.
