Hsc70 is a novel interactor of NF-kappaB p65 in living hippocampal neurons.

Hsc70 is a novel interactor of NF-kappaB p65 in living hippocampal neurons.
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DOI:
10.1371/journal.pone.0065280
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Kaltschmidt C
Kaltschmidt C
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Klenke C;Widera D;Engelen T;Müller J;Noll T;Niehaus K;Schmitz ML;Kaltschmidt B;Kaltschmidt C

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在神经元中通过NF-κB的信号传导依赖于与相互作用物如动力蛋白/动力肌动蛋白运动复合物的复合物形成,并且可以由突触激活触发。然而,到目前为止,神经元NF-κB的详细相互作用图是缺失的。在这项研究中,我们使用质谱法来鉴定脑内NF-κB p65的新相互作用物。Hsc 70是一种新的NF-κB p65神经元相互作用因子。在HEK 293细胞中,通过免疫共沉淀显示了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。用脱氧精胍菌素(DSG)阻断Hsc 70可显著降低谷氨酸刺激后神经元中NF-κB p65的核转位和报告基因检测显示的转录活性。此外,通过siRNA敲低Hsc 70显著降低神经元NF-κB活性。总之,这些数据为Hsc 70作为NF-κB p65的新型神经元相互作用物提供了证据。
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.
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