Willmann Katharina L, Sacco Roberto, Martins Rui, Garncarz Wojciech, Krolo Ana, Knapp Sylvia, Bennett Keiryn L, Boztug Kaan
CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, 1090 Vienna, Austria.
Department of Medicine I, Laboratory of Infection Biology, Medical University of Vienna, 1090 Vienna, Austria.
J Proteome Res. 2016 Sep 2;15(9):2900-2909. doi: 10.1021/acs.jproteome.5b01004. Epub 2016 Aug 1.
NF-κB signaling is a central pathway of immunity and integrates signal transduction upon a wide array of inflammatory stimuli. Noncanonical NF-κB signaling is activated by a small subset of TNF family receptors and characterized by NF-κB2/p52 transcriptional activity. The medical relevance of this pathway has recently re-emerged from the discovery of primary immunodeficiency patients that have loss-of-function mutations in the MAP3K14 gene encoding NIK. Nevertheless, knowledge of protein interactions that regulate noncanonical NF-κB signaling is sparse. Here we report a detailed state-of-the-art mass spectrometry-based protein-protein interaction network including the noncanonical NF-κB signaling nodes TRAF2, TRAF3, IKKα, NIK, and NF-κB2/p100. The value of the data set was confirmed by the identification of interactions already known to regulate this pathway. In addition, a remarkable number of novel interactors were identified. We provide validation of the novel NIK and IKKα interactor FKBP8, which may regulate processes downstream of noncanonical NF-κB signaling. To understand perturbed noncanonical NF-κB signaling in the context of misregulated NIK in disease, we also provide a differential interactome of NIK mutants that cause immunodeficiency. Altogether, this data set not only provides critical insight into how protein-protein interactions can regulate immune signaling but also offers a novel resource on noncanonical NF-κB signaling.
核因子κB（NF-κB）信号传导是免疫的核心途径，可整合多种炎症刺激下的信号转导。非经典NF-κB信号传导由一小部分肿瘤坏死因子（TNF）家族受体激活，并以NF-κB2/p52转录活性为特征。最近，通过发现编码NF-κB诱导激酶（NIK）的丝裂原活化蛋白激酶激酶激酶14（MAP3K14）基因存在功能丧失突变的原发性免疫缺陷患者，该信号传导途径的医学相关性再次受到关注。然而，关于调节非经典NF-κB信号传导的蛋白质相互作用的了解仍然很少。在此，我们报告了一个基于质谱的详细的最新蛋白质-蛋白质相互作用网络，其中包括非经典NF-κB信号传导节点肿瘤坏死因子受体相关因子2（TRAF2）、肿瘤坏死因子受体相关因子3（TRAF3）、IκB激酶α（IKKα）、NIK和NF-κB2/p100。通过鉴定已知调节该信号传导途径的相互作用，证实了该数据集的价值。此外，还鉴定出了大量新的相互作用蛋白。我们验证了新的NIK和IKKα相互作用蛋白FK506结合蛋白8（FKBP8），它可能调节非经典NF-κB信号传导的下游过程。为了了解在疾病中NIK调节异常情况下非经典NF-κB信号传导的紊乱，我们还提供了导致免疫缺陷的NIK突变体的差异相互作用组。总之，该数据集不仅为蛋白质-蛋白质相互作用如何调节免疫信号传导提供了关键见解，还为非经典NF-κB信号传导提供了新的资源。
