14-3-3 epsilon dynamically interacts with key components of mitogen-activated protein kinase signal module for selective modulation of the TNF-alpha-induced time course-dependent NF-kappaB activity.
14-3-3 epsilon dynamically interacts with key components of mitogen-activated protein kinase signal module for selective modulation of the TNF-alpha-induced time course-dependent NF-kappaB activity.
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DOI：
10.1021/pr9011377
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发表时间：
2010-06
影响因子：
4.4
通讯作者：
Shuai Zuo;Yan Xue;Siwei Tang;Jun Yao;Ruyun Du;Pengyuan Yang;Xian Chen
中科院分区：
文献类型：
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作者：
Shuai Zuo;Yan Xue;Siwei Tang;Jun Yao;Ruyun Du;Pengyuan Yang;Xian Chen
炎症受到核因子-κ B（NF-κ B）的严格调节，如果不加抑制，用于细胞因子过度产生的NF-κ B过度活化可导致包括致癌在内的各种致病后果。一种促炎细胞因子，肿瘤坏死因子-α（TNF-α），可用于探索可能的机制，其中未知的功能途径调节NF-κ B活性调节TNF-α诱导的炎症。鉴于14-3-3家族蛋白的多功能性质以及最近发现它们存在于TNF-α/NF-κ B通路网络中，我们使用双标记定量蛋白质组学方法首先分析TNF-α诱导的14-3-3 β相互作用伴侣，14-3-3 β是家族中特征最少的14-3-3异构体。我们首次发现，TNF-α刺激增强了14-3-3 β与丝裂原活化蛋白激酶（MAPK）信号模块中的一些关键组分之间的相互作用，该信号模块位于NF-κ B的紧邻上游，包括转化生长因子-β活化激酶-1（TAK 1）及其相互作用蛋白，蛋白磷酸酶2C β（PPM 1B）。通过使用共聚焦激光扫描，我们观察到TNF-α诱导的14-3-3 β，TAK 1和蛋白磷酸酶2C β（PPM 1B）之间的共定位，并且这些相互作用在不同的细胞类型中也是TNF-α诱导的。此外，我们发现在对TNF-α的细胞应答的整个过程中，14-3-3 β和这两种蛋白质之间的相互作用是动态的，并且与NF-κ B活性的时程依赖性变化密切相关，这表明这些14-3-3 β相互作用是TNF-α信号传导调节NF-κ B活性的关键点。然后，我们假设了一个机制的观点，描述了14-3-3 α如何协调其与TAK 1和PPM 1B的动态相互作用，以差异调节TNF-α诱导的NF-κ B活性变化。通过使用生物信息学工具，我们构建了包含我们蛋白质组学研究中鉴定的大多数14-3-3 β相互作用蛋白的网络。我们发现，14-3-3 β协调MAPK信号模块和其他分子途径/生物过程之间的交叉对话，主要包括蛋白质代谢和合成，DNA修复和细胞周期调节，其中可以系统地定位治疗干预的药理学靶点。
Inflammation is tightly regulated by nuclear factor-kappa B (NF-kappaB), and if left unchecked excessive NF-kappaB activation for cytokine overproduction can lead to various pathogenic consequences including carcinogenesis. A proinflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha), can be used to explore possible mechanisms whereby unknown functional pathways modulate the NF-kappaB activity for regulating TNF-alpha-induced inflammation. Given the multifunctional nature of 14-3-3 family proteins and the recent finding of their presence in the TNF-alpha/NF-kappaB pathway network, we used a dual-tagging quantitative proteomic method to first profile the TNF-alpha-inducible interacting partners of 14-3-3 epsilon, the least characterized 14-3-3 isomer in the family. For the first time, we found that TNF-alpha stimulation enhances the interactions between 14-3-3 epsilon and some key components in the mitogen-activated protein kinase (MAPK) signal module which is located at the immediate upstream of NF-kappaB, including transforming growth factor-beta activated kinase-1 (TAK1) and its interacting protein, protein phosphatase 2C beta (PPM1B). By using confocal laser scanning, we observed the TNF-alpha-induced colocalizations among 14-3-3 epsilon, TAK1, and protein phosphatase 2C beta (PPM1B), and these interactions were also TNF-alpha-inducible in different cell types. Further, we found that during the full course of the cellular response to TNF-alpha, the interactions between 14-3-3 epsilon and these two proteins were dynamic and were closely correlated with the time course-dependent changes in NF-kappaB activity, suggesting that these 14-3-3 epsilon interactions are the critical points of convergence for TNF-alpha signaling for modulating NF-kappaB activity. We then postulated a mechanistic view describing how 14-3-3 epsilon coordinates its dynamic interactions with TAK1 and PPM1B for differentially modulating TNF-alpha-induced changes in NF-kappaB activity. By using bioinformatics tools, we constructed the network involving most of the 14-3-3 epsilon interacting proteins identified in our proteomic study. We revealed that 14-3-3 epsilon coordinates the cross talks between the MAPK signal module and other molecular pathways/biological processes primarily including protein metabolism and synthesis, DNA repair, and cell cycle regulation where pharmacological targets for therapeutic intervention could be systematically located.