Novel Circadian Mutant of Drosophila

描述（由申请人提供）：许多行为、细胞和生化过程是由内源性生物钟控制的，这些自我维持的生物钟背后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，因此了解它们的基本机制可以为现代医学的许多领域做出贡献。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，这种振荡在翻译后水平上由几种激酶的活性控制，包括DBT酪蛋白激酶i。本应用旨在阐明DBT调节PER振荡的机制。第一个目标是确定与DBT和PER相互作用的其他蛋白质，以调节DBT的活性，并带来DBT磷酸化PER的生化后果。这个目标将使用果蝇细胞培养系（S2细胞）和成年果蝇头。目的2将确定在与成年蝇头昼夜节律功能相关的各种条件下，PER中被DBT和其他激酶磷酸化的位点。基于质谱的蛋白质组学方法将是目标1和目标2的主要焦点。最后，Aim 3将讨论蛋白质/蛋白质相互作用和特定位点与PER的磷酸化如何调节涉及PER和DBT的过程。因此，目标3将解决目标1和目标2中确定的相互作用和磷酸化位点的生物学后果。

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.