Autoinflammatory mutation in NLRC4 reveals a leucine-rich repeat (LRR)–LRR oligomerization interface
| Autor: | Heike Schützle, Reinhard Berner, Kai Lehmberg, Dominic De Nardo, Sebastian Brenner, Sigrun R. Hofmann, Huasong Zeng, Bingtai Lu, Stefan Winkler, Fiona Moghaddas, Ping Zeng, Li Zhang, Ian P. Wicks, Suyun Cheng, Yuanbo Zhao, Yuxia Zhang, Peter E. Czabotar, Christian M. Hedrich, Xiaoyun Chen, Scott W. Canna, Seth L. Masters |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Male NAIP NLR family apoptosis inhibitor protein Inflammasomes THP-1 Cells Mutant CARD Caspase activation and recruitment domain medicine.disease_cause NLRC4 NOD-like receptor family CARD-containing 4 protein CRISPR Clustered Regularly Interspaced Short Palindromic Repeats NLRC4 NLRC4-AID NLRC4-associated autoinflammatory disorder Immunology and Allergy Mutation HD Hinge domain NOD-like receptor Inflammasome ASC Apoptosis-associated Speck-like protein containing a caspase recruitment domain Syndrome LRR Leucine-rich repeat Cell biology NLRP3 NOD-like receptor family pyrin domain containing 3 NBD Nucleotide-binding domain Cyclic nucleotide-binding domain Female macrophage activation syndrome IL-18 medicine.drug Autoinflammatory disease periodic fever syndrome WT Wild-type Immunology Leucine-rich repeat Biology NOD Nucleotide-binding oligomerization domain Article 03 medical and health sciences Protein Domains inflammasome Leucine medicine AIFEC Autoinflammation with infantile enterocolitis Humans Nod-like receptor IPAF CSF Cerebrospinal fluid Calcium-Binding Proteins Infant Newborn KO Knockout Infant IL-18BP IL-18 binding protein Macrophage Activation CARD Signaling Adaptor Proteins 030104 developmental biology HEK293 Cells MAS Macrophage activation syndrome T3SS Type 3 secretion system Inflammasome complex |
| Zdroj: | The Journal of Allergy and Clinical Immunology JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY |
| ISSN: | 1097-6825 0091-6749 |
| Popis: | Background Monogenic autoinflammatory disorders are characterized by dysregulation of the innate immune system, for example by gain-of-function mutations in inflammasome-forming proteins, such as NOD-like receptor family CARD-containing 4 protein (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 2 unrelated patients with early-onset macrophage activation syndrome harboring the same de novo mutation in NLRC4. In vitro inflammasome complex formation was quantified by using flow cytometric analysis of apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) specks. Clustered Regularly Interspaced Short Palindromic Repeats (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-1β/IL-18 were quantified by using flow cytometry and ELISA, respectively. Results The p.W655C NLRC4 mutation caused increased ASC speck formation, caspase-1–dependent cell death, and IL-1β/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 2 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 type 3 secretion system 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 and previously unrecognized role in oligomerization of the NLRC4 inflammasome complex. Graphical abstract |
| Databáze: | OpenAIRE |
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