Molecular and Cellular Studies of Circadian Rhythms

描述(申请人提供)：我们一直在以果蝇为模型系统研究昼夜行为节律的分子控制。最初以苍蝇为特征的基因的同源基因，现在已经与控制包括鱼、青蛙、老鼠和人类在内的脊椎动物的节律行为和生理有关。我们建议对果蝇和哺乳动物的时钟中的两种激酶GSK-3和DBT进行研究。我们的一些初步工作提出了一种研究某些人类抑郁障碍的新方法。我们工作的其他方面可以从组织昼夜行为和生理反应的时钟中识别新的输出路径。我们将讨论以下问题：(1)GSK3是否像在果蝇中一样，在哺乳动物的发条过程中发挥核心作用？锂和丙戊酸是治疗双相情感障碍的首选药物，它们是如何阻止生物钟的？(2)GSK-3是否通过磷酸化TIM来影响飞行时钟？PER/TIM核转位是否需要GSK-3活性？(3)TIM是否在哺乳动物的生物钟中起作用？(4)DBT的体外活性是什么？(5)DBT依赖的磷酸化是否调节PER的核积累？PER的磷酸化如何影响TIM的磷酸化？(6)DBT与PER的物理联系是否将该激酶的其他底物置于昼夜节律控制之下？

DESCRIPTION (provided by applicant): We have been studying the molecular control of circadian behavioral rhythms using Drosophila as a model system. Homologues 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. We propose investigations of two kinases, GSK-3 and DBT, in the clocks of Drosophila and mammals. Some of our preliminary work suggests a novel approach to the study of certain human depressive disorders. Other aspects of our work could identify new output pathways from the clock that organize circadian behavioral and physiological responses. The following will be addressed: (1) Does GSK3 have a central role in the mammalian clockworks as it does in Drosophila? How do lithium and valproic acid, preferred agents for managing bipolar disorder, stop the circadian clock? (2) Does GSK-3 affect the fly clock by phosphorylating TIM? Is GSK-3 activity required for PER/TIM nuclear translocation? (3) Does TIM have a role in the mammalian clock? (4) What are the in vitro activities of DBT? (5) Does DBT-dependent phosphorylation regulate nuclear accumulation of PER? How does PER phosphorylation influence phosphorylation of TIM? (6) Does DBT's physical association with PER put other substrates of the kinase under circadian control?