Post-transcriptional regulations of proteomes in stress and senescence

项目总结 转录后机制在蛋白质水平调控基因表达方面起着基础性作用， 并经常与应激反应、衰老和疾病有关。这个项目的目标是开发 并应用多种组学方法来研究调节蛋白质的转录后机制 多个组织的组成及其对蛋白抑制应激源的反应能力。在最近的工作中，我们团队 已经开发出了旨在阐明蛋白质异构体的质谱学和多组学方法 组成和时空动力学。在这些进展的基础上，我们将在这里重点关注 三种已知在应激反应中影响蛋白质翻译的转录后机制。 具体地说，AIM 1将整合蛋白质组学和转录组学数据，以确定替代方案的作用 剪接在调节主要异构体丰度、产生替代蛋白形式和影响蛋白质中的作用 在哺乳动物组织中的定位。目标2将确定差异表达、定位和靶点 包括百草枯和阿霉素在内的蛋白质平衡应激反应中的RNA结合蛋白 和过氧化氢在体外。最后，《目标3》将考察翻译的结构和相互作用 包括核心核糖体和越来越多的已知核糖体相关蛋白的装置， 它们已经成为微调单个转录的翻译效率的重要因素 以及相关的蛋白质合成率。 拟议中的实验将询问转录后调控之间的关系 和应力响应，同时生成包括等形式解析的新的数据集， 正常、应激和衰老/衰老蛋白质组的时空图谱。我们预计结果将是 导致对应激反应和弹性的基本细胞过程的新见解，这些过程将与 对多种系统的研究。

PROJECT SUMMARY Post-transcriptional mechanisms play a fundamental role in regulating gene expression at the protein level, and are frequently implicated in stress response, aging, and diseases. The goal of this project is to develop and apply multi-omics methods to examine the post-transcriptional mechanisms that regulate protein composition of multiple tissues and their ability to respond to proteostatic stressors. In recent work, our team has developed mass spectrometry and multi-omics methods that are designed to elucidate the protein isoform composition and spatiotemporal dynamics. Building on these progresses, we will focus here on the roles of three post-transcriptional mechanisms known to influence protein translation in stress response. Specifically, Aim 1 will integrate proteomics and transcriptomics data to identify the role of alternative splicing in modulating principal isoform abundance, creating alternative proteoforms, and influencing protein localization in mammalian tissues. Aim 2 will determine the differential expression, localization, and targets of RNA-binding proteins in proteostatic stress responses including paraquat in vivo as well as doxorubicin and hydrogen peroxide in vitro. Finally, Aim 3 will examine the configuration and interactome of the translation apparatus including the core ribosome and an increasing number of known ribosome-associated proteins, which have emerged as important factors that can fine-tune the translational efficiency of individual transcripts and the associated protein synthesis rates. The proposed experiments will interrogate the relationships between post-transcriptional regulation and stress response, and at the same time generate novel data sets including isoform-resolved, spatiotemporal atlases of the normal, stressed, and aged/senescent proteomes. We anticipate the results will lead to novel insights into basic cellular processes of stress response and resilience that will be relevant to studies of multiple systems.