Wagner Sebastian A, Satpathy Shankha, Beli Petra, Choudhary Chunaram
Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark Department of Medicine, Hematology/Oncology, Goethe University School of Medicine, Frankfurt, Germany German Cancer Consortium (DKTK), Heidelberg, Germany German Cancer Research Center (DKFZ), Heidelberg, Germany
Department of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
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.
肿瘤坏死因子-α（TNF-α）是先天性免疫和促炎反应的关键调节因子。然而，TNF-α受体相关信号复合物（TNF-RSC）的组成以及下游信号网络的结构尚未完全明确。我们采用定量质谱法证明，TNF-α刺激可诱导广泛的蛋白质磷酸化，且磷酸化范围呈时间依赖性扩展。TNF-α刺激还可诱导TNF-RSC组分的快速泛素化。对TNF-RSC组成的时间分析确定SPATA2为TNF-RSC的一个新组分。SPATA2 N端预测的泛素结合蛋白（PUB）结构域与CYLD的泛素特异性蛋白酶（USP）结构域相互作用，而SPATA2的C端与HOIP相互作用。SPATA2是CYLD募集至TNF-RSC所必需的。SPATA2的下调增强核因子-κB（NF-κB）的转录激活，并抑制TNF-α诱导的坏死性凋亡，表明SPATA2在调节TNF-α信号转导结果中具有重要作用。综上所述，我们的研究绘制了TNF-α信号转导的详细图谱，确定SPATA2为TNF-α信号转导的一个新组分，并为进一步的功能研究提供了丰富资源。
