Mammalian circadian clock: genetics of PERIOD complex composition and structure.

哺乳动物生物钟：周期复杂组成和结构的遗传学。

• 批准号: 9336364
• 负责人: CHARLES J WEITZ
• 金额: $ 56.7万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336364
• 关键词: ARNTL gene; Address; Automobile Driving; Behavior; Behavior Control; Behavioral; Biochemistry; Biological; Biological Pacemakers; Biological Process; Biological Rhythm; Brain; Cell Nucleus; Circadian Rhythms; Collaborations; Complement; Complex; Cryoelectron Microscopy; Cytoplasm; DNA Binding; Defect; Diabetes Mellitus; Diagnosis; Dimensions; Disease; Electron Microscopy; Eukaryota; Feedback; Feeding behaviors; Functional disorder; Gene Targeting; Genetic; Genetic Transcription; Genotype; Goals; Hand; Health; Health Hazards; Heart; Human; Human Genetics; Image; Imagery; Individual; Investigation; Knowledge; Laboratories; Lead; Letters; Link; Liver; Mammals; Metabolic; Metabolic syndrome; Metabolism; Molecular; Mus; Mutant Strains Mice; Negative Staining; Nuclear; Nuclear Protein; Nuclear Proteins; Nuclear Structure; Obesity; Occupational Health; Peripheral; Phenotype; Physiological; Physiology; Play; Preparation; Property; Protein Analysis; Proteins; Resolution; Role; Sleep Disorders; Sleep Wake Cycle; Structure; System; Tissues; Transcription Regulatory Protein; Vitronectin; Work; circadian pacemaker; feeding; global health; improved; insight; microscopic imaging; mutant; particle; programs; protein complex; protein function; protein structure; sleep regulation; structural biology; success; three dimensional structure; transcription factor; virtual

Project Summary Circadian clocks are endogenous oscillators that drive daily rhythms of biological processes. In mammals, circadian clocks are found in the brain and in most peripheral tissues. Together, the distributed clocks constitute the fundamental timing system that coordinates daily behavior, physiology, and metabolism. The mammalian clock is built from a transcriptional feedback loop that generates circadian rhythms at the molecular level. The three PERIOD (PER) and two CRYPTOCHROME (CRY) proteins, transcriptional autoregulatory proteins that are dedicated clock components, form a large nuclear protein complex (PER complex) that lies at the heart of the feedback loop. In recent years our laboratory has used preparative purification to analyze the constituent proteins of the nuclear PER complex, providing new mechanistic insights into its transcriptional actions. In the present application we begin to address the question of how the nuclear PER complex works in an integrated fashion as a macromolecular machine, a challenging but essential next step in understanding. Building on our successful efforts, we propose to characterize the properties, composition, and structural organization of PER complexes from wildtype and mutant mice lacking individual PER or CRY proteins. By uniting preparative purification of PER complexes with mouse genetics and structural biology, the application aims to determine how each PER and CRY protein contributes to the assembly, composition, and three- dimensional structure of the nuclear PER complex. If successful, the project offers to deepen our knowledge of the clock mechanism substantially, possibly to an atomic level of resolution. Advances in understanding the mammalian circadian clock will have important implications for our view of the genetic control of behavior and physiology, as well as for human health and disease. Studies from mouse genetics, human genetics, and occupational health indicate that defects of clock function lead to broad behavioral and metabolic dysfunction, producing, for example, disrupted sleep-wake cycles, abnormal feeding, and a metabolic syndrome closely resembling early-stage diabetes. If successful, the proposed study will provide new insights into genetic and molecular mechanisms controlling fundamental behavioral and physiological programs linked to major diseases.

项目摘要 昼夜节律钟是驱动生物钟的每日节律的内源性振荡器。 流程.在哺乳动物中，生物钟存在于大脑和大多数外周组织中。 这些分布式时钟共同构成了协调日常工作的基本计时系统 行为、生理和新陈代谢。 哺乳动物的生物钟是建立在一个转录反馈回路上的， 分子水平的节奏。三个周期（PER）和两个周期（CRY） 蛋白质，转录自动调节蛋白，是专门的时钟组件，形成一个 大核蛋白复合物（PER复合物），位于反馈环的中心。近几 多年来，我们的实验室一直使用制备纯化来分析蛋白质的组成。 核PER复合物，提供了新的机制的见解，其转录行动。 在本申请中，我们开始解决核PER如何 作为一种高分子机器，一种具有挑战性的， 理解的重要下一步。在我们成功努力的基础上，我们建议 表征PER复合物的性质、组成和结构组织， 缺乏个别PER或CRY蛋白的野生型和突变型小鼠。通过联合准备 本申请利用小鼠遗传学和结构生物学纯化PER复合物， 确定每一个PER和CRY蛋白如何有助于组装，组成，和三个- 核PER复合物的三维结构。如果成功，该项目将深化我们的 时钟机制的基本知识，可能达到原子级的分辨率。 了解哺乳动物生物钟的进展将具有重要意义 我们对行为和生理的遗传控制的看法，以及对人类健康和 疾病来自小鼠遗传学、人类遗传学和职业健康的研究表明， 生物钟功能的缺陷会导致广泛的行为和代谢功能障碍， 例如，睡眠-觉醒周期中断，喂养异常和代谢综合征密切相关 类似于早期糖尿病如果成功，拟议的研究将提供新的见解， 控制基本行为和生理的遗传和分子机制 与重大疾病有关的项目。