title: SPATA2 Links CYLD to LUBAC, Activates CYLD, and Controls LUBAC Signaling.
abstract: The linear ubiquitin chain assembly complex (LUBAC) regulates immune signaling, and its function is regulated by the deubiquitinases OTULIN and CYLD, which associate with the catalytic subunit HOIP. However, the mechanism through which CYLD interacts with HOIP is unclear. We here show that CYLD interacts with HOIP via spermatogenesis-associated protein 2 (SPATA2). SPATA2 interacts with CYLD through its non-canonical PUB domain, which binds the catalytic CYLD USP domain in a CYLD B-box-dependent manner. Significantly, SPATA2 binding activates CYLD-mediated hydrolysis of ubiquitin chains. SPATA2 also harbors a conserved PUB-interacting motif that selectively docks into the HOIP PUB domain. In cells, SPATA2 is recruited to the TNF receptor 1 signaling complex and is required for CYLD recruitment. Loss of SPATA2 increases ubiquitination of LUBAC substrates and results in enhanced NOD2 signaling. Our data reveal SPATA2 as a high-affinity binding partner of CYLD and HOIP, and a regulatory component of LUBAC-mediated NF-κB signaling.
authors: Division of Protein and Nucleic Acid Chemistry, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
authors: Paul R
authors: PR
authors: Elliott
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK.
authors: Derek
authors: D
authors: Leske
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK.
authors: Matous
authors: M
authors: Hrdinka
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK.
authors: Katrin
authors: K
authors: Bagola
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK.
authors: Berthe K
authors: BK
authors: Fiil
authors: Division of Protein and Nucleic Acid Chemistry, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
authors: Stephen H
authors: SH
authors: McLaughlin
authors: Division of Protein and Nucleic Acid Chemistry, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
authors: Jane
authors: J
authors: Wagstaff
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK.
authors: Norbert
authors: N
authors: Volkmar
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK.
authors: John C
authors: JC
authors: Christianson
authors: TDI Mass Spectrometry Laboratory, Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7FZ, UK.
authors: Benedikt M
authors: BM
authors: Kessler
authors: Division of Protein and Nucleic Acid Chemistry, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
authors: Stefan M V
authors: SM
authors: Freund
authors: Division of Protein and Nucleic Acid Chemistry, MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK. Electronic address: dk@mrc-lmb.cam.ac.uk.
authors: David
authors: D
authors: Komander
authors: Ludwig Institute for Cancer Research, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Oxford OX3 7DQ, UK. Electronic address: mads.gyrd-hansen@ludwig.ox.ac.uk.
authors: Mads
authors: M
authors: Gyrd-Hansen
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D000074749:Deubiquitinating Enzyme CYLD; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation; D006801:Humans; D007113:Immunity, Innate; D007700:Kinetics; D062105:Molecular Docking Simulation; D016328:NF-kappa B / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D053473:Nod2 Signaling Adaptor Protein / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013379:Substrate Specificity; D025521:Tumor Suppressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D025801:Ubiquitin / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry* / Q000235:genetics / Q000276:immunology
chemical_list: D016328:NF-kappa B; C429018:NOD2 protein, human; D053473:Nod2 Signaling Adaptor Protein; D011506:Proteins; D011994:Recombinant Proteins; C425523:SPATA2 protein, human; D025521:Tumor Suppressor Proteins; D025801:Ubiquitin; C558338:RNF31 protein, human; D044767:Ubiquitin-Protein Ligases; D010450:Endopeptidases; C582107:OTULIN protein, human; C409984:CYLD protein, human; D000074749:Deubiquitinating Enzyme CYLD