Non-transcriptional regulation of circadian physiology

昼夜节律生理学的非转录调节

• 批准号: 10406109
• 负责人: JOANNA Chungyen CHIU
• 金额: $ 8.56万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-16 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10406109
• 关键词: Address; Animals; Behavior; Behavioral Assay; Bypass; Cell physiology; Clock protein; Conflict (Psychology); Cues; Drosophila genus; Eating; Equilibrium; Event; Exhibits; Fasting; Gene Expression; Genetic Transcription; Goals; Human; Label; Life; Life Style; Light; Link; Metabolic; Metabolic Diseases; Metabolism; Modeling; Modernization; Molecular; Nutrient; Organism; Periodicity; Phosphorylation; Physiology; Play; Post-Translational Protein Processing; Proteins; Proteomics; Regulation; Role; Schedule; Signal Transduction; Testing; Time; Time-restricted feeding; Work; circadian; circadian pacemaker; circadian regulation; feeding; metabolomics; new therapeutic target; protein function; time use; transcription factor

PROJECT SUMMARY Circadian clocks are endogenous protein machines that integrate external time cues and internal metabolic states to regulate daily rhythms in physiology and behavior in organisms from all kingdoms of life. In the natural world, environmental zeitgebers enable the animal circadian clock to control timing of food intake. Nutrient influx can therefore provide metabolic signals to reinforce environmental signals, promoting synchrony in circadian physiology to balance metabolism and energy use. Initial efforts to dissect the underpinnings of the circadian oscillator and its control over rhythms of life focused on regulation at the transcriptional level, as the core oscillator proteins are transcription factors that collaborate to govern rhythmic expression of genes involved in diverse cellular processes. More recent studies have uncovered complementary non- transcriptional mechanisms, including protein post-translational modifications (PTMs), that are critical for circadian timekeeping. The overall goal of this project is to understand the mechanisms by which metabolic and environmental signals integrate at the post-translational level to regulate circadian physiology, and more importantly the consequences when these signals that have evolved to cooperate are in conflict. We will use the diurnal Drosophila model to test the central hypothesis that nutrient influx through clock-controlled feeding activity regulates the interplay between phosphorylation and O-linked N-Acetylglucosaminylation (O- GlcNAcylation) of cellular proteins to modulate time-of-day specific functions. Protein O- GlcNAcylation is highly sensitive to metabolic input and may play a dominant role in extensive remodeling of cellular protein functions, bypassing changes in gene expression. In Aim 1, we will use time-restricted feeding (TRF) in combination with targeted metabolomics and chemoenzymatic O-GlcNAc labeling to establish the relationships between feeding-fasting cycle, nutrient influx, and O-GlcNAcylation status of cellular proteins. In Aim 2, we will identify cellular proteins that exhibit daily interplay between O-GlcNAcylation and phosphorylation using label- free proteomic approaches. In Aim 3, we will characterize the function of clock protein O- GlcNAcylation events by utilizing tried-and-true molecular and Drosophila behavioral assays. By addressing the 3 questions: When, What, and Why, we will advance our understanding on metabolic regulation of circadian physiology via post-translational mechanisms. This project will have broad significance as cross-talk between protein phosphorylation and O-GlcNAcylation is extensive and modulates a wide range of cellular processes. Our findings may identify new therapeutic targets to alleviate circadian and metabolic disorders.

项目概要 生物钟是内源性蛋白质机器，它整合了外部时间线索和 内部代谢状态调节生物体的生理和行为的日常节律 一切生命王国。在自然界中，环境时间使动物的昼夜节律得以实现 时钟来控制食物摄入的时间。因此，营养流入可以提供代谢信号 强化环境信号，促进昼夜生理同步以达到平衡 新陈代谢和能量利用。剖析昼夜节律振荡器基础的初步努力 而其对生命节律的控制则集中在转录水平的调控上，为核心 振荡蛋白是协作控制基因节律表达的转录因子 参与多种细胞过程。最近的研究发现互补的非 转录机制，包括蛋白质翻译后修饰 (PTM) 对于昼夜节律计时至关重要。该项目的总体目标是了解 代谢和环境信号在翻译后整合的机制 调节昼夜节律生理的水平，更重要的是，当这些 演化而来的合作信号是相互冲突的。我们将使用昼夜果蝇模型 检验营养物质通过时钟控制的摄食活动流入的中心假设 调节磷酸化和 O-连接的 N-乙酰葡糖胺酰化 (O- GlcNAcyllation）细胞蛋白质来调节一天中不同时间的特定功能。蛋白O- GlcNAc 酰化对代谢输入高度敏感，可能在广泛的代谢过程中发挥主导作用。 重塑细胞蛋白质功能，绕过基因表达的变化。在目标 1 中，我们将 将限时喂养 (TRF) 与靶向代谢组学相结合 化学酶 O-GlcNAc 标记可建立进食-禁食周期之间的关系， 营养流入和细胞蛋白质的 O-GlcNAc 酰化状态。在目标 2 中，我们将识别细胞 使用标记显示 O-GlcNAcNA 酰化和磷酸化之间日常相互作用的蛋白质 免费蛋白质组学方法。在目标 3 中，我们将描述时钟蛋白 O-的功能 利用经过验证的分子和果蝇行为测定进行 GlcNAc 酰化事件。经过 解决三个问题：何时、什么和为什么，我们将加深对以下问题的理解： 通过翻译后机制调节昼夜节律生理学。该项目将 具有广泛的意义，因为蛋白质磷酸化和 O-GlcNAcNA 酰化之间的串扰是 广泛并调节广泛的细胞过程。我们的发现可能会发现新的 缓解昼夜节律和代谢紊乱的治疗目标。