Cell Biology, University of Bielefeld, Bielefeld, Germany.
PLoS One. 2013 Jun 7;8(6):e65280. doi: 10.1371/journal.pone.0065280. Print 2013.
Signaling via NF-κB in neurons depends on complex formation with interactors such as dynein/dynactin motor complex and can be triggered by synaptic activation. However, so far a detailed interaction map for the neuronal NF-κB is missing. In this study we used mass spectrometry to identify novel interactors of NF-κB p65 within the brain. Hsc70 was identified as a novel neuronal interactor of NF-κB p65. In HEK293 cells, a direct physical interaction was shown by co-immunoprecipitation and verified via in situ proximity ligation in healthy rat neurons. Pharmacological blockade of Hsc70 by deoxyspergualin (DSG) strongly decreased nuclear translocation of NF-κB p65 and transcriptional activity shown by reporter gene assays in neurons after stimulation with glutamate. In addition, knock down of Hsc70 via siRNA significantly reduced neuronal NF-κB activity. Taken together these data provide evidence for Hsc70 as a novel neuronal interactor of NF-κB p65.
神经元中的 NF-κB 信号转导依赖于与动力蛋白/动力蛋白复合物等相互作用子的复杂形成，并且可以被突触激活所触发。然而，到目前为止，神经元 NF-κB 的详细相互作用图谱仍然缺失。在这项研究中，我们使用质谱法鉴定了大脑中 NF-κB p65 的新相互作用子。热休克蛋白 70（Hsc70）被鉴定为 NF-κB p65 的新型神经元相互作用子。在 HEK293 细胞中，通过共免疫沉淀显示出直接的物理相互作用，并通过健康大鼠神经元中的原位邻近连接进行了验证。用去氧精胍（DSG）抑制 Hsc70 的药理学阻断作用，在谷氨酸刺激后的神经元中，NF-κB p65 的核易位和报告基因分析显示的转录活性显著降低。此外，通过 siRNA 敲低 Hsc70 显著降低了神经元 NF-κB 活性。综上所述，这些数据为 Hsc70 作为 NF-κB p65 的新型神经元相互作用子提供了证据。
