Collaborative Research: Biochemical Basis of Cellular Circadian Behavior

合作研究：细胞昼夜节律行为的生化基础

• 批准号: 1656647
• 负责人: Andrew Liu
• 金额: $ 64.3万
• 依托单位: University of Memphis
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1656647&HistoricalAwards=false
• 关键词: Collaborative; Research; Biochemical; Basis; Cellular

Non-Technical Paragraph By establishing the genetic basis of rhythmic behavior, circadian clock research is a great success story in neuroscience. The molecular clock consists of positive and negative components that form a feedback loop that underlies brain rhythms. However, "the whole is greater than the sum of its parts," and we do not yet know how a handful of components coordinate to give rise to emergent daily rhythms. Therefore, this research aims to identify the biochemical properties of core clock proteins that enable rhythmic behavior. To do this, the team has developed an integrative approach that combines cell-based genetics and continuous monitoring of circadian behavior with biochemical/biophysical methods. This approach will allow them to study the function of clock proteins not only in test tubes in isolation, but also in the context of oscillatory behavior in interacting cells. This work will reveal how the "gears and springs" of the clock interact and function together, and this knowledge will increase our understanding of how the brain keeps time, as well as how proteins interact to regulate complex brain functions. The PI and his colleagues will teach a lecture/lab course that uses a "from math to genes to behavior" platform and provides cross-disciplinary training for graduate students. The team will also teach real world concepts in the local community about the health implications of circadian rhythms, which relates to people of all backgrounds and ages. These broader impacts will strengthen the nation's scientific infrastructure and improve people's health awareness. Technical Paragraph Genetic studies have identified several core clock components that form a negative feedback mechanism, which underlies circadian behavior. It is well established that, in the core feedback loop, BMAL1 and CLOCK are the two transcription activators and CRY serves as the chief repressor. However, negative feedback in transcription does not necessarily warrant recurring cellular processes with a ~24 hr periodicity. The circadian rhythm is an emergent property enabled by the core clock factors, but the biochemical basis of cellular circadian oscillation is not well understood. Recent studies from this research team show that the structurally flexible C-termini of BMAL1 and CLOCK play essential roles in regulating dynamic interactions with other clock factors to enable circadian oscillations. In the proposed research, the team will employ an integrated approach that combines cell-based genetics and kinetic bioluminescence assays with biochemical and biophysical methods to study how their C-termini use dynamic interactions to regulate rhythm amplitude and period length. In this way, protein function is assessed both in vitro and in the context of cellular circadian behavior. By providing a biochemical basis of cellular circadian behavior, this research will define the molecular architecture of the circadian transcription complex, advance current understanding of the negative feedback mechanism, and further the goals of behavioral neurobiology. During the research, the PI and his colleagues will provide cross-disciplinary education and training for graduate students and inform the general public about the important health implications of circadian rhythms.

非技术段落通过建立节奏行为的遗传基础，昼夜节律研究是神经科学中的巨大成功故事。分子时钟由构成脑节律基础的反馈回路的正组件和负组成。但是，“整体大于其各个部分的总和”，我们尚不知道少数组件如何协调产生新兴的每日节奏。因此，该研究旨在确定能够有节奏行为的核心时钟蛋白的生化特性。为此，该团队开发了一种综合方法，该方法将基于细胞的遗传学和对昼夜节律行为的持续监测与生化/生物物理方法相结合。这种方法将使他们能够研究时钟蛋白的功能，不仅是孤立的，而且还研究了相互作用细胞中振荡行为的情况。这项工作将揭示时钟的“齿轮和弹簧”如何相互作用和功能，并且这些知识将增加我们对大脑如何保持时间的理解，以及蛋白质如何相互作用以调节复杂的大脑功能。 PI和他的同事将教授讲课/实验室课程，该课程使用“从数学到基因到行为”平台，并为研究生提供跨学科培训。该团队还将在当地社区中教授现实世界的概念，了解与各个背景和年龄的人有关的昼夜节律的健康影响。这些更广泛的影响将增强国家的科学基础设施并提高人们的健康意识。技术段落遗传研究已经确定了形成负面反馈机制的几个核心时钟成分，这些反馈机制是昼夜节律行为的基础。众所周知，在核心反馈循环中，BMAL1和时钟是两个转录激活剂，而Cry则是主要阻遏物。但是，转录中的负反馈不一定保留约24小时周期性的经常性细胞过程。昼夜节律是由核心时钟因素启用的新兴特性，但是细胞昼夜节律振荡的生化基础尚不清楚。该研究小组的最新研究表明，BMAL1和时钟的结构灵活的C末端在调节与其他时钟因子的动态相互作用中起着至关重要的作用，以实现昼夜节律振荡。在拟议的研究中，该团队将采用一种综合方法，该方法将基于细胞的遗传学和动力学发光测定与生化和生物物理方法相结合，以研究其C-terni如何使用动态相互作用来调节节奏幅度和周期长度。这样，在体外和细胞昼夜行为的情况下都评估了蛋白质功能。通过提供细胞昼夜行为的生化基础，这项研究将定义昼夜节律转录复合物的分子结构，对当前对负面反馈机制的理解以及行为神经生物学的目标。在研究期间，PI及其同事将为研究生提供跨学科的教育和培训，并将昼夜节律的重要健康含义告知公众。