Interdisciplinary studies of the Drosophila Circadian Clock

果蝇昼夜节律钟的跨学科研究

• 批准号: 7354090
• 负责人: Michael Warren Young
• 金额: $ 36.92万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-03 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7354090
• 关键词: Affect; Amino Acids; Area; Behavior; Behavioral; Binding; Biochemical Genetics; Biological Models; Chloride Anion Exchanger; Circadian Rhythms; Class; Daily; Data; Depth; Development; Drosophila genus; Elements; Fishes; Flavins; Gene Expression; Gene Proteins; Genes; Genome; Glycogen Synthase Kinase 3; Head; Homologous Gene; Human; Individual; Interdisciplinary Study; Investigation; Lethal Genes; Life; Light; Link; Maps; Molecular; Mus; Mutation; Mutation Analysis; Orthologous Gene; Output; Periodicity; Phosphorylation; Physiological; Physiology; Protein Splicing; Proteins; Protocols documentation; Rana; Recovery; Reporting; Research; Role; SLC26A3 gene; Series; Structure; Techniques; Tissues; Vertebrates; Work; base; chemical genetics; circadian behavioral rhythms; circadian pacemaker; fly; insight; interest; novel; null mutation; programs; protein function; research study; response

We have been studying the molecular control of circadian behavioral rhythms using Drosophila as a model system. Orthologs of genes initially characterized in the fly,have now been linked to the control of rhythmic behavior and physiology in vertebrates, including fish, frogs, mice and humans. Here we propose three classes of interdisciplinary investigations of the Drosophila clock. (1) We will conduct collaborative structural studies that can help us determine how specific regulatory actions, previously recognized genetically and biochemically, are performed by certain domains of the PER protein. Does the structure of PER indicate how it interacts with TIM and DBT? Do such data suggest how TIM suppresses PER's phosphorylation by DBT? Does a putative LOV domain near the N-terminus of PER possess a flavin binding domain? (2) We will generate new mutations for the analysis of vital clock genes by Conditional Protein Splicing. Our studies of hyper- and hypo-morphic mutations of GSK-3, dbt, vri, and Pdp-1 indicate that each is a key component of the Drosophila clock. However, because null mutations of these genes are lethal, it has been difficult to determine their full effects on the clock. We are developing a form of chemical genetics in which small diffusible molecules will reversibly control the presence of each vital protein in living flies. (3) We will produce new microarray and statistical approaches to clarify features of rhythmic genome activity and to determine if flies use a single molecular mechanism to generate all circadian rhythms. To begin to link the clock to specific behavioral and physiological outputs, we will investigate a novel set of genes that are regulated by a paired action of light and the circadian clock.

我们一直在以果蝇为模型研究昼夜行为节律的分子控制 系统。最初在果蝇中鉴定的基因直向同源物现在已与节律的控制联系起来 脊椎动物（包括鱼、青蛙、小鼠和人类）的行为和生理学。这里我们提出三个 果蝇时钟的跨学科研究类别。 （一）开展协同结构 研究可以帮助我们确定先前认识到的基因和特定监管行为如何 生物化学上，这些过程是由 PER 蛋白的某些结构域执行的。 PER的结构是否表明如何 它与 TIM 和 DBT 相互作用吗？这些数据是否表明 TIM 如何通过 DBT 抑制 PER 磷酸化？ PER N 末端附近的推定 LOV 结构域是否具有黄素结合结构域？ (2) 我们将 通过条件蛋白质剪接产生新的突变，用于分析生命钟基因。我们的研究 GSK-3、dbt、vri 和 Pdp-1 的超态和亚态突变表明每一个都是 果蝇时钟。然而，由于这些基因的无效突变是致命的，因此很难 确定它们对时钟的全部影响。我们正在开发一种化学遗传学形式，其中小的 扩散分子将可逆地控制活果蝇中每种重要蛋白质的存在。 (3) 我们将生产 新的微阵列和统计方法来阐明节律基因组活性的特征并确定是否 果蝇使用单一分子机制来产生所有昼夜节律。开始将时钟链接到 特定的行为和生理输出，我们将研究一组新的基因，这些基因受 光和生物钟的配对作用。