Functional Translatome Proteomics Reveal Converging and Dose-Dependent Regulation by mTORC1 and eIF2α
Functional Translatome Proteomics Reveal Converging and Dose-Dependent Regulation by mTORC1 and eIF2α
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DOI：
10.1016/j.molcel.2019.11.010
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发表时间：
2020-02-20
期刊：
影响因子：
16
通讯作者：
Muench, Christian
中科院分区：
文献类型：
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作者：
Klann, Kevin;Tascher, Georg;Muench, Christian
翻译的调节在压力下是必不可少的。然而，由关键的翻译应激反应-整合应激反应（ISR）和mTORC1-调控的蛋白质的精确集合仍然难以捉摸。我们开发了多路增强蛋白质动力学（mePROD）蛋白质组学，将信号放大添加到动态SILAC和多路复用中，以测量蛋白质合成的急剧变化。用ISR/mTORC1调节应激物处理细胞，我们显示了广泛的翻译调节，类似于20%的蛋白质合成率高度降低。比较免疫缺陷亚蛋白质组揭示了广泛的重叠，表明靶特异性是在蛋白质水平上实现的，而不是通过途径激活。滴定帽依赖性翻译抑制证实，个别蛋白质的合成是由响应于全球翻译衰减的内在特性控制的。这项研究报告了一种高度敏感的方法来测量新生链水平的相对翻译，并深入了解了两种关键细胞途径ISR和mTORC 1如何调节翻译组以指导细胞在应激时的生存。
Regulation of translation is essential during stress. However, the precise sets of proteins regulated by the key translational stress responses-the integrated stress response (ISR) and mTORC1-remain elusive. We developed multiplexed enhanced protein dynamics (mePROD) proteomics, adding signal amplification to dynamic-SILAC and multiplexing, to enable measuring acute changes in protein synthesis. Treating cells with ISR/mTORC1-modulating stressors, we showedextensive translatomemodulation with similar to 20% of proteins synthesized at highly reduced rates. Comparing translation-deficient sub-proteomes revealed an extensive overlap demonstrating that target specificity is achieved on protein level and not by pathway activation. Titrating cap-dependent translation inhibition confirmed that synthesis of individual proteins is controlled by intrinsic properties responding to global translation attenuation. This study reports a highly sensitive method to measure relative translation at the nascent chain level and provides insight into howthe ISR and mTORC1, two key cellular pathways, regulate the translatome to guide cellular survival upon stress.