Investigating Circadian Post-Transcriptional Regulation.

研究昼夜节律转录后调节。

• 批准号: 10228665
• 负责人: Jennifer Hurley
• 金额: $ 53.26万
• 依托单位: RENSSELAER POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10228665
• 关键词: Address; Affect; Animal Model; Architecture; Bacteria; Behavior; Biochemical; Biochemical Genetics; Biological Clocks; Biological Models; Bread; Cardiovascular Diseases; Cell physiology; Circadian Dysregulation; Circadian Rhythms; Disease; Ensure; Feedback; Genes; Genetic Transcription; Goals; Hour; Human; Immune system; Investigation; Life Style; Link; Malignant Neoplasms; Maps; Medical; Mental disorders; Modeling; Modernization; Molds; Molecular Conformation; Mus; Nature; Neurospora crassa; Output; Phase; Physiological; Physiology; Play; Population; Post-Transcriptional Regulation; Proteins; Proteome; Regulation; Reproducibility; Risk; Role; Sleep; Time; Translations; Work; arm; base; circadian; circadian regulation; cost; fitness; flexibility; genetic manipulation; insight; luminescence; macrophage; molecular clock; novel; promoter; protein protein interaction; therapeutic effectiveness; transcriptome

Project Summary/Abstract: Circadian rhythms are highly conserved, roughly 24-hour, physiological cycles that adjust innumerable actions, affecting everything from luminescence in bacteria to sleep in humans. Through the ideal programming of behavior, it is believed that these rhythms enhance fitness by ensuring that many organismal functions are optimally synchronized with the appropriate phase of the circadian day. Disruption of proper circadian timing negatively impacts the human long-term medical outlook, making it critical to understand the mechanism underlying circadian regulation over cellular physiology. Circadian rhythms are controlled via a highly-regulated transcription-translation based negative feedback loop, or clock. The current paradigm for clock regulation over cellular physiology is that transcriptional activity from the positive arm of the transcription–translation negative feedback loop drives the expression of a host of gene promoters that modulate organismal behavior. However, mounting evidence suggests that circadian regulation is imparted on cellular physiology beyond the level of transcription and that the negative arm may play a role in this regulation. The long-term goal of our work is to determine the extent of this post-transcriptional regulation on cellular physiology and to identify the mechanistic underpinnings of circadian post-transcriptional regulation. As a mechanism for keeping time, transcription–translation negative feedback loops are highly conserved and much of what is understood about the molecular clock comes from the investigation of model systems. Therefore, we will exploit the simplicity and reproducibility of model systems to cost-effectively address our hypotheses. To determine the extent of circadian post-transcriptional regulation, we will analyze the transcriptome and proteome of murine macrophages over circadian time. As mice are a common model for the human immune system, our study will garner insights into both the extent of circadian post-transcriptional regulation as well as investigate clock regulation on the immune system. To tackle the mechanistic underpinnings of post-transcriptional regulation, we will utilize Neurospora crassa, a bread mold whose ease of biochemical and genetic manipulation is unparalleled in any other eukaryotic clock model system. We hypothesize that the negative arm may control circadian output via transient protein-protein interactions, which are synchronized by timed conformational changes that are enabled by the negative arm’s inherently flexible biochemical nature. We will create a Conformational/Temporal Interactome (CTI) map of circadian negative arm proteins to validate our hypothesis. Due to the conservation of clock architecture, the results of this work have the potential to define several novel and unrecognized paradigms in clock regulation over cellular physiology.

项目摘要/摘要： 昼夜节律是高度保守的，大约24小时的物理周期，可调节无数 动作，影响从细菌中的发光到人类睡眠的一切。通过理想的编程 在行为上，人们认为这些节奏通过确保许多有机功能是 与昼夜节日的适当阶段最佳同步。适当的昼夜节律时机 负面影响人类的长期医学前景，使了解机制至关重要 对细胞生理学的昼夜节律调节。昼夜节律通过高度调节 基于转录翻译的负反馈循环或时钟。时钟调节的当前范例 细胞生理是从转录 - 翻译阴性的正臂的转录活性 反馈回路驱动了调节有机行为的许多基因启动子的表达。然而， 越来越多的证据表明，昼夜节律的调节是超出细胞生理学的 转录和负臂可能在该法规中发挥作用。我们工作的长期目标是 确定此后转录后对细胞生理的调节的程度，并确定机理 昼夜节目后调节的基础。 作为保留时间的机制，转录 - 翻译负反馈循环是高度保守的 关于分子时钟的许多理解的内容来自模型系统的投资。 因此，我们将探讨模型系统的简单性和可重复性，以成本效益地解决我们的 假设。为了确定昼夜节目后调节的程度，我们将分析 在昼夜节律时间内，鼠巨噬细胞的转录组和蛋白质组。因为小鼠是一个常见模型 人类免疫系统，我们的研究将获得对昼夜节目后的范围的见解 调节以及调查免疫系统的时钟调节。解决机械基础 在转录后调节中，我们将利用神经孢子虫，一种面包模具，其易于生化 在任何其他真核时钟模型系统中，遗传操纵是无与伦比的。我们假设 负臂可以通过瞬时蛋白质蛋白质相互作用来控制昼夜节律的输出，这些相互作用由 负面臂的固有灵活的生化性质实现了定时的构象变化。我们 将创建昼夜节律负臂蛋白的构象/颞相互作用图（CTI）图来验证我们 假设。由于时钟体系结构的保护，这项工作的结果有可能定义 关于细胞生理的时钟调节中的几种新颖和未被认可的范例。