Department of Biochemistry, Faculty of Science, Masaryk University, Brno, Czech Republic.
Michael Smith Laboratories, University of British Columbia, Vancouver, Canada.
Mol Cell Proteomics. 2024 Apr;23(4):100744. doi: 10.1016/j.mcpro.2024.100744. Epub 2024 Feb 28.
NF-κB pathway is involved in inflammation; however, recent data shows its role also in cancer development and progression, including metastasis. To understand the role of NF-κB interactome dynamics in cancer, we study the complexity of breast cancer interactome in luminal A breast cancer model and its rearrangement associated with NF-κB modulation. Liquid chromatography-mass spectrometry measurement of 160 size-exclusion chromatography fractions identifies 5460 protein groups. Seven thousand five hundred sixty eight interactions among these proteins have been reconstructed by PrInCE algorithm, of which 2564 have been validated in independent datasets. NF-κB modulation leads to rearrangement of protein complexes involved in NF-κB signaling and immune response, cell cycle regulation, and DNA replication. Central NF-κB transcription regulator RELA co-elutes with interactors of NF-κB activator PRMT5, and these complexes are confirmed by AlphaPulldown prediction. A complementary immunoprecipitation experiment recapitulates RELA interactions with other NF-κB factors, associating NF-κB inhibition with lower binding of NF-κB activators to RELA. This study describes a network of pro-tumorigenic protein interactions and their rearrangement upon NF-κB inhibition with potential therapeutic implications in tumors with high NF-κB activity.
NF-κB 通路参与炎症；然而，最近的数据表明它在癌症的发展和进展中也有作用，包括转移。为了了解 NF-κB 相互作用组动力学在癌症中的作用，我们研究了腔 A 型乳腺癌模型中乳腺癌相互作用组的复杂性及其与 NF-κB 调节相关的重排。使用液相色谱-质谱法对 160 个排阻色谱法分馏物进行测量，鉴定出 5460 个蛋白质组。通过 PrInCE 算法重建了这些蛋白质之间的 7568 个相互作用，其中 2564 个相互作用已在独立数据集得到验证。NF-κB 调节导致参与 NF-κB 信号转导和免疫反应、细胞周期调节和 DNA 复制的蛋白质复合物的重排。NF-κB 的中央转录调节剂 RELA 与 NF-κB 激活剂 PRMT5 的相互作用体共洗脱，这些复合物通过 AlphaPulldown 预测得到证实。互补的免疫沉淀实验重现了 RELA 与其他 NF-κB 因子的相互作用，将 NF-κB 抑制与 NF-κB 激活剂与 RELA 的结合降低联系起来。这项研究描述了一个促进肿瘤发生的蛋白质相互作用网络及其在 NF-κB 抑制时的重排，这可能对 NF-κB 活性高的肿瘤具有治疗意义。
