Post-transcriptional regulations of proteomes in stress and senescence

项目摘要 本行政补充申请正在根据NOT-GM-22提交给PA-20-272- 017.父项目R01-GM144456的范围不会更改。请求的补充内容将替换为 关键硬件基础设施，以利用现代功能加速我们的研究，并允许 生产力。家长项目总结：转录后机制在调控中起着基础性作用 基因在蛋白质水平上的表达，经常与应激反应、衰老和疾病有关。这个 本项目的目标是开发和应用多种组学方法来检测转录后 调节多种组织的蛋白质组成及其对内源性和非特异性反应能力的机制 环境蛋白抑制应激源。在最近的工作中，我们的团队开发了质谱学和多光谱 旨在阐明蛋白质异构体组成和时空动力学的组学方法。 在这些进展的基础上，我们将在这里集中讨论已知的三种转录后机制的作用 影响应激反应中的蛋白质翻译。具体地说，AIM 1将整合蛋白质组学和 转录学数据以确定选择性剪接在调节主要异构体丰度中的作用， 创造另一种蛋白质形式，并影响跨组织(心、肺、肌肉、脑)的蛋白质定位， 性别和寿命。目标2将确定RNA结合的差异表达、定位和靶点 蛋白质在体内对包括百草枯在内的内源性或外源性应激源的应激反应以及 阿霉素和过氧化氢的体外实验。最后，《目标3》将研究 翻译装置包括核心核糖体和越来越多的已知核糖体- 相关蛋白，它们已经成为微调翻译效率的重要因素 单个转录本和相关的蛋白质合成率。拟议的实验将审问 转录后调控与应激反应之间的关系，并同时产生新的 数据集包括正常、应激和老龄化/衰老的等形式解析的时空地图集 蛋白质组。我们预计，这些结果将导致对应激反应的基本细胞过程的新见解 和恢复力，这将与多系统的研究相关。

Project Summary This administrative supplement application is being submitted to PA-20-272 in accordance with NOT-GM-22- 017. The scope of the parent project R01-GM144456 will not be changed. The requested supplement will replace critical hardware infrastructure to accelerate our research with modern capabilities and permit increased productivity. Parent 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 intrinsic and environmental 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 across tissues (heart, lung, muscle, brain), sexes, and lifespan. Aim 2 will determine the differential expression, localization, and targets of RNA-binding proteins in proteostatic stress responses to intrinsic or extrinsic stressors 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.