Abstract
Cpdm (chronic proliferative dermatitis) mice develop chronic dermatitis and an immunodeficiency with increased serum IgM, 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). Spontaneous null mutations in the Sharpin (SHANK-associated RH domain interacting protein in postsynaptic density) gene are responsible for the cpdm phenotype in mice. SHARPIN shows significant similarity to HOIL-1L (also known as RBCK1), a component of linear ubiquitin chain assembly complex (LUBAC), which induces NF-κB activation through conjugation of linear polyubiquitin chains to NEMO. 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.
MeSH Terms
Animals
CD40 Ligand/metabolism
Carrier Proteins/metabolism
Cells, Cultured
HEK293 Cells
Humans
Intracellular Signaling Peptides and Proteins/metabolism
Mice
Multiprotein Complexes/chemistry,metabolism
NF-kappa B/metabolism
Nerve Tissue Proteins/deficiency,genetics,metabolism
Tumor Necrosis Factor-alpha/metabolism
Ubiquitin/metabolism
Ubiquitin-Protein Ligase Complexes/chemistry,metabolism
Ubiquitin-Protein Ligases/metabolism
Chemicals
Carrier Proteins
HOIL-1L protein, mouse
Intracellular Signaling Peptides and Proteins
Multiprotein Complexes
NEMO protein, mouse
NF-kappa B
Nerve Tissue Proteins
Tumor Necrosis Factor-alpha
Ubiquitin
sharpin
CD40 Ligand
Ubiquitin-Protein Ligase Complexes
Rnf31 protein, mouse
Ubiquitin-Protein Ligases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Tokunaga Fuminori
Department of Biophysics and Biochemistry, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan.
Nakagawa Tomoko
Nakahara Masaki
Saeki Yasushi
Taniguchi Masami
Sakata Shin-ichi
Tanaka Keiji
Nakano Hiroyasu
Iwai Kazuhiro
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