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.

项目摘要 该项目在NIGMS资助的两个研究项目的基础上，追求两个研究方向。两者都有 项目是松散联系的，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。酵母菌模型的前期工作 已经建立了不同的泛素介导的信号事件，这些信号事件传递新陈代谢和 环境状态对细胞周期机制的影响。对主宰概念的分子理解 因此，泛素信号转导是本申请第一部分的主题。项目1研究酵母菌和 哺乳动物细胞系模型来定义泛素信号的一般概念。这些实验建立在 我们开发了过多的工具来分析泛素信号和泛素的生化和生理学 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)泛素连接酶如何 以上下文特定的方式识别底物？(4)信号是如何通过磷酸化实现的 泛素。项目1中提议的工作将定义泛素信号转导方面的详细分子洞察力 蛋白水解性和非蛋白水解性。 Project-2专注于哺乳动物系统，我们是否发现了重要的连接体 蛋氨酸代谢与细胞增殖。其中包括对蛋白磷酸酶2A(PP2A)的调节。 以及选定转录本的RNA CAP甲基化。在哺乳动物细胞中对PP2A的研究将集中在这一角色上 PP2A催化亚单位羧基末端甲基化作为蛋氨酸传感器的研究 新陈代谢。实验将扩展到确定几个PP2A相互作用的蛋白质组图谱 首选去甲基化的PP2A复合体。我们将仔细分析这些交互并定义 它们在向细胞周期机制传递新陈代谢状态方面的作用。信使核糖核酸CAP的相关研究 甲基化将扩展我们最近的发现，即一小群RNA对微妙的东西高度敏感 由蛋氨酸代谢控制的细胞甲基化潜能的波动。这些 RNA在其信使核糖核酸帽上发生低甲基化，因此当蛋氨酸 是有限的。其目标是了解使某些mRNA对波动高度敏感的机制。 在蛋氨酸代谢中，并发现与细胞周期控制的机制联系。 理解将蛋氨酸代谢与细胞增殖承诺相结合的分子概念 新的治疗策略，特别是癌症和其他与年龄相关的疾病的治疗。因此， 这项提议旨在发展分子洞察力，以了解一种到目前为止在分子上尚未探索的基础， 具有巨大治疗潜力的生物过程。