SPATA2 Links CYLD to LUBAC, Activates CYLD, and Controls LUBAC Signaling

Mol Cell. 2016 Sep 15;63(6):990-1005. doi: 10.1016/j.molcel.2016.08.001. Epub 2016 Aug 30.

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.

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Cloning, Molecular
  • Crystallography, X-Ray
  • Deubiquitinating Enzyme CYLD
  • Endopeptidases / chemistry
  • Endopeptidases / genetics
  • Endopeptidases / immunology
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Gene Expression Regulation
  • Humans
  • Immunity, Innate
  • Kinetics
  • Molecular Docking Simulation
  • NF-kappa B / chemistry*
  • NF-kappa B / genetics
  • NF-kappa B / immunology
  • Nod2 Signaling Adaptor Protein / chemistry
  • Nod2 Signaling Adaptor Protein / genetics
  • Nod2 Signaling Adaptor Protein / immunology
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Structure, Secondary
  • Proteins / chemistry*
  • Proteins / genetics
  • Proteins / immunology
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • Substrate Specificity
  • Tumor Suppressor Proteins / chemistry*
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / immunology
  • Ubiquitin / chemistry*
  • Ubiquitin / genetics
  • Ubiquitin / immunology
  • Ubiquitin-Protein Ligases / chemistry*
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / immunology

Substances

  • NF-kappa B
  • NOD2 protein, human
  • Nod2 Signaling Adaptor Protein
  • Proteins
  • Recombinant Proteins
  • SPATA2 protein, human
  • Tumor Suppressor Proteins
  • Ubiquitin
  • RNF31 protein, human
  • Ubiquitin-Protein Ligases
  • Endopeptidases
  • gumby protein, human
  • CYLD protein, human
  • Deubiquitinating Enzyme CYLD