Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, INSERM, CNRS, Marseille, France.
Centre d'Immunophénomique, Aix Marseille Université, INSERM, CNRS UMR, Marseille, France.
Front Immunol. 2023 Mar 15;14:1139123. doi: 10.3389/fimmu.2023.1139123. eCollection 2023.
The propagation and diversification of signals downstream of the T cell receptor (TCR) involve several adaptor proteins that control the assembly of multimolecular signaling complexes (signalosomes). The global characterization of changes in protein-protein interactions (PPI) following genetic perturbations is critical to understand the resulting phenotypes. Here, by combining genome editing techniques in T cells and interactomics studies based on affinity purification coupled to mass spectrometry (AP-MS) analysis, we determined and quantified the molecular reorganization of the SLP76 interactome resulting from the ablation of each of the three GRB2-family adaptors. Our data showed that the absence of GADS or GRB2 induces a major remodeling of the PPI network associated with SLP76 following TCR engagement. Unexpectedly, this PPI network rewiring minimally affects proximal molecular events of the TCR signaling pathway. Nevertheless, during prolonged TCR stimulation, GRB2- and GADS-deficient cells displayed a reduced level of activation and cytokine secretion capacity. Using the canonical SLP76 signalosome, this analysis highlights the plasticity of PPI networks and their reorganization following specific genetic perturbations.
T 细胞受体 (TCR) 下游信号的传播和多样化涉及几种衔接蛋白，这些蛋白控制着多分子信号复合物（信号小体）的组装。遗传扰动后蛋白质-蛋白质相互作用 (PPI) 变化的全面表征对于理解所产生的表型至关重要。在这里，我们通过结合 T 细胞中的基因组编辑技术和基于亲和纯化结合质谱分析的互作组学研究，确定并量化了 SLP76 互作组在三个 GRB2 家族衔接蛋白缺失后的分子重排。我们的数据表明，GADS 或 GRB2 的缺失会导致 TCR 结合后与 SLP76 相关的 PPI 网络发生重大重塑。出乎意料的是，这种 PPI 网络重排对 TCR 信号通路的近端分子事件影响很小。然而，在长期的 TCR 刺激下，GRB2 和 GADS 缺陷细胞表现出较低的激活水平和细胞因子分泌能力。使用经典的 SLP76 信号小体，该分析强调了 PPI 网络的可塑性及其在特定遗传扰动后的重组。
