Autoinflammatory mutation in NLRC4 reveals an LRR-LRR oligomerization interface

Moghaddas, F; Zeng, P; Zhang, Y; Schutzle, H; Brenner, S; Hofmann, SR; Berner, R; Zhao, Y; Lu, B; Chen, X; Zhang, L; Cheng, S; Winkler, S; Lehmberg, K; Canna, SW; Czabotar, PE; Wicks, IP; De Nardo, D; Hedrich, CM; Zeng, H; Masters, SL

Author(s): Moghaddas, F; Zeng, P; Zhang, Y; Schutzle, H; Brenner, S; Hofmann, SR; Berner, R; Zhao, Y; Lu, B; Chen, X; Zhang, L; Cheng, S; Winkler, S; Lehmberg, K; Canna, SW; Czabotar, PE; Wicks, IP; De Nardo, D; Hedrich, CM; Zeng, H; Masters, SL;
Details: Publication Year 2018-05-17,Volume 142,Issue #6,Page 1956-1967e6
Journal Title: Journal of Allergy and Clinical Immunology
Publication Type: Journal Article
Abstract: BACKGROUND: Monogenic autoinflammatory disorders are characterised by dysregulation of the innate immune system, for example by gain-of-function mutations in inflammasome forming proteins such as NLRC4. OBJECTIVE: Here we investigate the mechanism by which a novel mutation in the leucine rich repeat (LRR) domain of NLRC4 (c.G1965C, p.W655C) contributes to autoinflammatory disease. METHODS: We studied two unrelated patients with early onset macrophage activation syndrome (MAS) harboring the same de novo mutation in NLRC4. In vitro, inflammasome complex formation was quantified using flow cytometric analysis of ASC specks. CRISPR/Cas9 techniques and lentiviral transduction were used to generate THP-1 cells with either wild-type or mutant NLRC4 cDNA. Cell death and release of IL-1beta/IL-18 were quantified by flow cytometry and ELISA respectively. RESULTS: The c.G1965C/p.W655C NLRC4 mutation caused increased ASC speck formation, caspase-1-dependent cell death, and IL-1beta/IL-18 production. ASC contributed to p.W655C NLRC4 mediated cytokine release, but not cell death. Mutation of p.W655 activated the NLRC4 inflammasome complex by engaging with two interfaces on the opposing LRR domain of the oligomer. One key set of residues (p.D1010, p.D1011, p.L1012 and p.I1015) participated in LRR-LRR oligomerization when triggered by mutant NLRC4 or T3SS effector (PrgI) stimulation of the NLRC4 inflammasome complex. CONCLUSION: This is the first report of a mutation in the LRR domain of NLRC4 causing autoinflammatory disease. c.G1965C/p.W655C NLRC4 increased inflammasome activation in vitro. Data generated from various NLRC4 mutations provides evidence that the LRR-LRR interface has an important, previously unrecognized role in oligomerization of the NLRC4 inflammasome complex.
Publisher: Elsevier
Research Division(s): Chemical Biology; Inflammation
PubMed ID: 29778503
Publisher's Version: https://doi.org/10.1016/j.jaci.2018.04.033
Open Access at Publisher's Site: https://doi.org/10.1016/j.jaci.2018.04.033
NHMRC Grants: NHMRC/1113577, NHMRC/1099262,
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2018-05-31 09:17:47

Last Modified: 2020-06-17 09:04:42