Analysis of clock proteins in their non-circadian roles

分析时钟蛋白的非昼夜节律作用

• 批准号: 7048732
• 负责人: Jadwiga M Giebultowicz
• 金额: $ 26.25万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7048732
• 关键词: Drosophilidae; arthropod genetics; biological clocks; biological signal transduction; circadian rhythms; developmental genetics; fertility; gene mutation; head; immunocytochemistry; immunoprecipitation; mass spectrometry; metabolism; nutrition; oogenesis; ovary; protein protein interaction; protein purification; protein structure function; Drosophilidae; arthropod genetics; biological clocks; biological signal transduction; circadian rhythms; developmental genetics; fertility; gene mutation; head; immunocytochemistry; immunoprecipitation; mass spectrometry; metabolism; nutrition; oogenesis; ovary; protein protein interaction; protein purification; protein structure function

DESCRIPTION (provided by applicant): Circadian clocks are evolutionary conserved coordinators of behavioral and physiological processes. Malfunctions of circadian clocks in humans lead to serious pathologies such as sleep disorders and cancer. Circadian timekeeping is accomplished by molecular feedback loops that involve several clock genes and their proteins. The role of two genes period (per) and timeless (tim) has been well established in the clock feedback loop, which operates in the model organism Drosophila melanogaster. The products of these two genes, proteins PER and TIM, translocate to cell nuclei and are subsequently degraded; both events are essential for clock function. Surprisingly, in the ovary, these proteins behave differently. Their levels do not cycle; instead, they remain stable and cytoplasmic at all times. We have evidence that non-circadian expression of PER and TIM in the ovary may have important functions in the modulation of egg production. We hypothesize that clock genes may be interacting with components of signaling pathways that govern metabolic homeostasis and nutrient allocation. We propose to use biochemical and genetic tools to test this hypothesis in two specific aims. First, we will study genetic and biochemical interactions of PER and TIM in the ovary and test their functional significance using fecundity related phenotypes. Second, we will perform protein interaction screens to identify novel proteins that may interact with cytoplasmic PER and TIM. Results obtained in this study will give us important insights into the functional significance of non-circadian expression of clock proteins. There is increasing evidence that genes that were thought to act exclusively as clock components have other important pleiotropic roles. They act in a non-circadian manner in both fly and mammals. Therefore, understanding the non-circadian functions of clock genes in a model organism should provide valuable insights into similar processes related to human health.

描述（由申请人提供）：昼夜节律是行为和生理过程的进化保守协调员。人类昼夜节律的故障导致严重的病理，例如睡眠障碍和癌症。昼夜节律是通过涉及多个时钟基因及其蛋白质的分子反馈回路来完成的。两个基因周期（per）和永恒（tim）的作用在时钟反馈回路中已经建立了，该反馈循环在模型有机体果蝇中运行。这两个基因的产物，蛋白质和TIM的蛋白质，转移到细胞核，随后降解。这两个事件对于时钟功能都是必不可少的。 令人惊讶的是，在卵巢中，这些蛋白质的行为不同。他们的水平不循环；相反，它们始终保持稳定和细胞质。我们有证据表明，在卵巢中，非circadian的表达在卵子的调节中可能具有重要功能。我们假设时钟基因可能正在与控制代谢稳态和营养分配的信号通路的组成部分相互作用。我们建议使用生化和遗传工具以两个具体目标来检验这一假设。首先，我们将研究卵巢中Per和Tim的遗传和生化相互作用，并使用与繁殖力相关的表型测试其功能意义。其次，我们将执行蛋白质相互作用筛选，以鉴定可能与细胞质per和tim相互作用的新型蛋白质。在这项研究中获得的结果将使我们对钟蛋白的非circadian表达的功能意义有重要见解。有越来越多的证据表明，人们被认为仅作为时钟成分具有其他重要的多效性作用的基因。他们在苍蝇和哺乳动物中都以非circ派的方式起作用。因此，了解模型生物体中时钟基因的非circadian功能应为与人类健康相关的类似过程提供有价值的见解。