title: CARD8 and NLRP1 undergo autoproteolytic processing through a ZU5-like domain.
abstract: The "Function to Find Domain" (FIIND)-containing proteins CARD8 (Cardinal; Tucan) and NLRP1 (NALP1; NAC) are well known components of inflammasomes, multiprotein complexes responsible for activation of caspase-1, a regulator of inflammation and innate immunity. Although identified many years ago, the role of the FIIND is unknown. Here, we report that CARD8 and NLRP1 undergo autoproteolytic cleavage at a conserved SF/S motif within the FIIND. Using bioinformatics and computational modeling approaches, we detected striking structural similarity between the FIIND and the ZU5-UPA domain present in the autoproteolytic protein PIDD. This allowed us to generate a three-dimensional model and to gain insights in the molecular mechanism of the cleavage. Site-directed mutagenesis experiments revealed that the second serine of the SF/S motif is required for CARD8 and NLRP1 autoproteolysis. Furthermore, we discovered an important function for conserved glutamic acid and histidine residues, located in proximity of the cleavage site in regulating the autoprocessing efficiency. Altogether, these results identify a function for the FIIND and show that CARD8 and NLRP1 are ZU5-UPA domain-containing autoproteolytic proteins, thus suggesting a novel mechanism for regulating innate immune responses.
authors.affiliation: Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Andrea
authors.initials: A
authors.lastname: D'Osualdo
authors.forename: Christian X
authors.initials: CX
authors.lastname: Weichenberger
authors.forename: Roland N
authors.initials: RN
authors.lastname: Wagner
authors.forename: Adam
authors.initials: A
authors.lastname: Godzik
authors.forename: John
authors.initials: J
authors.lastname: Wooley
authors.forename: John C
authors.initials: JC
authors.lastname: Reed
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D001665:Binding Sites; D053468:CARD Signaling Adaptor Proteins / Q000378:metabolism*; D002460:Cell Line; D006801:Humans; D007113:Immunity, Innate; D058847:Inflammasomes; D046912:Multiprotein Complexes / Q000276:immunology; D000070576:NLR Proteins; D009363:Neoplasm Proteins / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism*; D017434:Protein Structure, Tertiary
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D053468:CARD Signaling Adaptor Proteins; C455623:CARD8 protein, human; D058847:Inflammasomes; D046912:Multiprotein Complexes; D000070576:NLR Proteins; C427903:NLRP1 protein, human; D009363:Neoplasm Proteins; D010447:Peptide Hydrolases
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