Leib Lisa, Juli Jana, Jurida Liane, Mayr-Buro Christin, Priester Jasmin, Weiser Hendrik, Wirth Stefanie, Hanel Simon, Heylmann Daniel, Weber Axel, Schmitz M Lienhard, Papantonis Argyris, Bartkuhn Marek, Wilhelm Jochen, Linne Uwe, Meier-Soelch Johanna, Kracht Michael
Rudolf Buchheim Institute of Pharmacology, Justus Liebig University, Giessen, Germany.
Institute of Biochemistry, Justus Liebig University, Giessen, Germany.
EMBO Rep. 2025 Feb;26(4):1144-1183. doi: 10.1038/s44319-024-00339-8. Epub 2025 Jan 3.
The protein interactome of p65/RELA, the most active subunit of the transcription factor (TF) NF-κB, has not been previously determined in living cells. Using p65-miniTurbo fusion proteins and biotin tagging, we identify >350 RELA interactors from untreated and IL-1α-stimulated cells, including many TFs (47% of all interactors) and >50 epigenetic regulators belonging to different classes of chromatin remodeling complexes. A comparison with the interactomes of two point mutants of p65 reveals that the interactions primarily require intact dimerization rather than DNA-binding properties. A targeted RNAi screen for 38 interactors and subsequent functional transcriptome and bioinformatics studies identify gene regulatory (sub)networks, each controlled by RELA in combination with one of the TFs ZBTB5, GLIS2, TFE3/TFEB, or S100A8/A9. The large, dynamic and versatile high-resolution interactome of RELA and its gene regulatory logics provides a rich resource and a new framework for explaining how RELA cooperativity determines gene expression patterns.
转录因子（TF）NF-κB最具活性的亚基p65/RELA的蛋白质相互作用组，此前尚未在活细胞中确定。利用p65-微型Turbo融合蛋白和生物素标记，我们从未经处理和IL-1α刺激的细胞中鉴定出超过350个RELA相互作用蛋白，包括许多转录因子（占所有相互作用蛋白的47%）以及超过50个属于不同染色质重塑复合体类别的表观遗传调节因子。与p65的两个点突变体的相互作用组进行比较发现，这些相互作用主要需要完整的二聚化，而非DNA结合特性。针对38个相互作用蛋白进行的靶向RNA干扰筛选以及后续的功能转录组和生物信息学研究，确定了基因调控（子）网络，每个网络由RELA与TFs ZBTB5、GLIS2、TFE3/TFEB或S100A8/A9之一共同控制。RELA庞大、动态且通用的高分辨率相互作用组及其基因调控逻辑，为解释RELA协同作用如何决定基因表达模式提供了丰富资源和新框架。
