SPATA2 links CYLD to the TNF-α receptor signaling complex and modulates the receptor signaling outcomes

EMBO J. 2016 Sep 1;35(17):1868-84. doi: 10.15252/embj.201694300. Epub 2016 Jun 15.


TNF-α is a key regulator of innate immune and proinflammatory responses. However, the composition of the TNF-α receptor-associated signaling complexes (TNF-RSC) and the architecture of the downstream signaling networks are incompletely understood. We employed quantitative mass spectrometry to demonstrate that TNF-α stimulation induces widespread protein phosphorylation and that the scope of phosphorylation expands in a temporal manner. TNF-α stimulation also induces rapid ubiquitylation of components of the TNF-RSC Temporal analysis of the TNF-RSC composition identified SPATA2 as a novel component of the TNF-RSC The predicted PUB domain in the N-terminus of SPATA2 interacts with the USP domain of CYLD, whereas the C-terminus of SPATA2 interacts with HOIP SPATA2 is required for recruitment of CYLD to the TNF-RSC Downregulation of SPATA2 augments transcriptional activation of NF-κB and inhibits TNF-α-induced necroptosis, pointing to an important function of SPATA2 in modulating the outcomes of TNF-α signaling. Taken together, our study draws a detailed map of TNF-α signaling, identifies SPATA2 as a novel component of TNF-α signaling, and provides a rich resource for further functional investigations.

Keywords: CYLD; LUBAC; SPATA2; TNF‐RSC; necroptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Deubiquitinating Enzyme CYLD
  • Humans
  • Mass Spectrometry
  • Phosphorylation
  • Protein Interaction Mapping
  • Protein Processing, Post-Translational
  • Proteins / metabolism*
  • Proteome / analysis
  • Receptors, Tumor Necrosis Factor / metabolism*
  • Signal Transduction*
  • Tumor Necrosis Factor-alpha / metabolism*
  • Tumor Suppressor Proteins / metabolism*
  • Ubiquitin-Protein Ligases / metabolism
  • Ubiquitination


  • Proteins
  • Proteome
  • Receptors, Tumor Necrosis Factor
  • SPATA2 protein, human
  • Tumor Necrosis Factor-alpha
  • Tumor Suppressor Proteins
  • RNF31 protein, human
  • Ubiquitin-Protein Ligases
  • CYLD protein, human
  • Deubiquitinating Enzyme CYLD