Novel Circadian Mutant of Drosophila

描述（由申请人提供）：许多行为、细胞和生化过程由内源性生物钟控制，这些自我维持的生物钟的分子机制是临床医生和基础科学家感兴趣的。昼夜节律的临床相关性在其对睡眠/觉醒周期的控制中最为明显，但许多其他医学相关过程也受到昼夜节律的调节。内分泌功能、癌症调节、精神疾病和药物代谢都受到生物钟的影响，因此了解其基本机制有助于现代医学的许多领域。在哺乳动物和果蝇中，这种分子机制的核心是PER蛋白的振荡，其在翻译后水平上由几种激酶（包括DBT酪蛋白激酶I）的活性控制。本申请旨在阐明DBT调节PER振荡的机制。第一个目标是鉴定与DBT和PER相互作用以调节DBT活性并引起DBT磷酸化PER的生化后果的其他蛋白质。这一目标将采用果蝇细胞培养系（S2细胞）以及成年果蝇头。目的2将确定的网站内的PER磷酸化的DBT和其他激酶，在各种条件下相关的昼夜节律功能的成年苍蝇头部。基于质谱的蛋白质组学方法将是目标1和2的主要重点。最后，目标3将解决蛋白质/蛋白质相互作用和磷酸化在特定位点与PER调节过程涉及PER和DBT。因此，目标3将解决目标1和2中确定的相互作用和磷酸化位点的生物学后果。 公共卫生相关性：昼夜节律是行为和生理的日常周期，例如我们的睡眠/觉醒行为，它们影响药物疗效，癌症，代谢和心理障碍。这个应用程序旨在了解产生生物钟的关键生化机制之一。该机制非常适合药物治疗，因此理解可能会影响许多人类疾病的治疗。

DESCRIPTION (provided by applicant): Many behavioral, cellular and biochemical processes are controlled by an endogenous circadian clock, and the molecular mechanisms underlying these self-sustaining circadian clocks are of interest to both clinicians and basic scientists. The clinical relevance of circadian rhythms is most obvious in their control of sleep/wake cycles, but many other medically relevant processes are also regulated by circadian rhythms. Endocrine function, cancer regulation, psychiatric disorders and drug metabolism are all affected by circadian clocks, and therefore knowledge of their basic mechanism can contribute to many areas of modern medicine. At the core of this molecular mechanism in mammals and flies are oscillations of the PER protein, which are controlled at the post-translational level by the activities of several kinases, including the DBT casein kinase I. This application proposes to elucidate the mechanism by which DBT regulates the oscillations of PER. The first aim will identify other proteins which interact with DBT and PER to regulate the activity of DBT and bring about the biochemical consequences of PER's phosphorylation by DBT. This aim will employ both a Drosophila cell culture line (S2 cells) as well as adult fly heads. Aim 2 will identify the sites within PER that are phosphorylated by DBT and other kinases, under various conditions relevant to circadian function in the adult fly head. A mass-spectrometry-based proteomic approach will be a major focus of both Aims 1 and 2. Finally, Aim 3 will address how protein/protein interactions and phosphorylation at particular sites with PER regulate processes involving PER and DBT. Aim 3 will therefore address the biological consequences of the interactions and phosphorylation sites identified in Aims 1 and 2. PUBLIC HEALTH RELEVANCE: Circadian rhythms are daily cycles of behavior and physiology, as exemplified by our sleep/wake behavior, and they affect drug efficacy, cancer, metabolism and psychological disorders. This application seeks to understand one of the key biochemical mechanisms that produce the circadian clock. The mechanism is very amenable to treatment with drugs, so the understanding may affect treatments of a number of human diseases.