Ubiquitin and Metabolite Signaling

泛素和代谢信号传导

• 批准号: 10552304
• 负责人: Peter Kaiser
• 金额: $ 44.98万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552304
• 关键词: Affect; Animals; Biochemistry; Biological Process; Catalytic Domain; Cell Cycle; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cell physiology; Cells; Communication; Complex; Cues; Development; Disease; Event; F-Box Proteins; Funding; Goals; Link; Longevity; Mammalian Cell; Mediating; Messenger RNA; Metabolic; Methionine; Methionine Metabolism Pathway; Methylation; Modeling; Molecular; National Institute of General Medical Sciences; Organism; Pathway interactions; Phosphorylation; Physiology; Protein phosphatase; Proteomics; RNA; RNA Caps; Reader; Regulation; Research; Role; Signal Transduction; System; Therapeutic; Transcript; Translating; Ubiquitin; Work; Yeast Model System; Yeasts; age related; cancer cell; cancer therapy; demethylation; experimental study; insight; mRNA capping; novel therapeutic intervention; preference; programs; response; sensor; tool; transmission process; treatment strategy; ubiquitin ligase

Project Summary This project pursues two research directions that build on two NIGMS funded research programs. Both projects are loosely connected and the pathways studied will help understand how metabolic and environmental cues are sensed and transmitted to the cell cycle machinery. Previous work in yeast models has established different ubiquitin-mediated signaling events that communicate metabolic and environmental states to the cell cycle machinery. Molecular understanding of the concepts that govern ubiquitin signaling are thus the topic of the first part of this application. Project-1 studies both yeast and mammalian cell line models to define general concepts of ubiquitin signaling. These experiments build on a plethora of tools we have developed to analyze biochemistry and physiology of ubiquitin signaling and will address the following questions: (1) How do readers of the ubiquitin signal distinguish different chain types? (2) How do F-box proteins sense metabolic and environmental states? (3) How do ubiquitin ligases recognize substrates in a context specific manner? (4) How is signaling achieved by phosphorylated ubiquitin. Proposed work in project 1 will define detailed molecular insight in aspects of ubiquitin signaling both proteolytic and non-proteolytic. Project-2 is focused on the mammalian system, were we discovered important connectors between methionine metabolism and cell proliferation. These include regulation of protein phosphatase 2A (PP2A) and RNA CAP methylation of selected transcripts. Work on PP2A in mammalian cells will focus on the role of methylation of the carboxy terminus of the catalytic subunit of PP2A as a sensor of methionine metabolism. Experiments will expand on proteomic profiling that identified several PP2A interaction partners with preference for the demethylated PP2A complex. We will dissect these interactions and define their role in communicating metabolic states to the cell cycle machinery. Studies related to mRNA CAP methylation will extend our recent findings that a small group of RNAs is highly sensitive to subtle fluctuations in the cellular methylation potential, which is controlled by methionine metabolism. These RNAs become hypomethylated on their mRNA CAPs, and thus are inefficiently translated when methionine is limiting. The goal is to understand mechanisms that make certain mRNAs hypersensitive to fluctuation in methionine metabolism, and to discover the mechanistic link to cell cycle control. Understanding the molecular concepts that integrate methionine metabolism with cell proliferation promise new therapeutic strategies, especially for the treatment of cancer and other age-related disorders. Thus, this proposal aims to development molecular insight into a fundamental, so far molecularly unexplored, biological process with great therapeutic potential.

项目概要 该项目以两个 NIGMS 资助的研究项目为基础，追求两个研究方向。两个都 项目之间的联系松散，所研究的途径将有助于理解新陈代谢和 环境信号被感知并传输到细胞周期机器。之前在酵母模型中的工作 已经建立了不同的泛素介导的信号传导事件，这些信号传导代谢和 环境状态对细胞周期机制的影响。对主导概念的分子理解 因此，泛素信号传导是本申请第一部分的主题。 Project-1 研究酵母和 哺乳动物细胞系模型来定义泛素信号传导的一般概念。这些实验建立在 我们开发了大量工具来分析泛素信号传导的生物化学和生理学 将解决以下问题：（1）泛素信号的阅读器如何区分不同的链 类型？ (2) F-box蛋白如何感知代谢和环境状态？ (3) 泛素连接酶是如何进行的 以上下文特定的方式识别底物？ (4) 磷酸化信号是如何实现的 泛素。项目 1 中拟议的工作将定义泛素信号传导方面的详细分子见解 分为蛋白水解的和非蛋白水解的。 Project-2 的重点是哺乳动物系统，我们是否发现了哺乳动物系统之间的重要连接器？ 蛋氨酸代谢和细胞增殖。其中包括蛋白磷酸酶 2A (PP2A) 的调节 以及所选转录本的 RNA CAP 甲基化。哺乳动物细胞中 PP2A 的研究重点是其作用 PP2A催化亚基羧基末端甲基化作为蛋氨酸传感器的研究 代谢。实验将扩展蛋白质组学分析，确定多种 PP2A 相互作用 偏好去甲基化 PP2A 复合物的合作伙伴。我们将剖析这些相互作用并定义 它们在将代谢状态传递给细胞周期机制方面发挥着作用。 mRNA CAP相关研究 甲基化将扩展我们最近的发现，即一小群 RNA 对细微的变化高度敏感。 细胞甲基化电位的波动，由蛋氨酸代谢控制。这些 RNA 在其 mRNA CAP 上变得低甲基化，因此当蛋氨酸存在时翻译效率低下 是有限的。目标是了解使某些 mRNA 对波动高度敏感的机制 蛋氨酸代谢，并发现与细胞周期控制的机制联系。 了解将蛋氨酸代谢与细胞增殖前景相结合的分子概念 新的治疗策略，特别是用于治疗癌症和其他与年龄相关的疾病。因此， 该提案旨在将分子洞察力发展为迄今为止分子尚未探索的基本原理， 具有巨大治疗潜力的生物过程。