Dissection of EARLY FLOWERING3 Function in the Arabidopsis Circadian Clock

描述（由申请人提供）：称为生物钟的内源性分子振荡器控制着从人类日常睡眠/觉醒周期到植物叶片运动的重要生物过程。生物钟的主要功能是精确协调生物体对日常环境变化的生理反应。在许多生物体中，环境刺激和生物钟整合的分子机制还没有得到很好的理解。我的目标是了解环境信号是如何整合到生物钟中的，以及参与这种整合的特定基因的功能，以拟南芥为模型系统。本提案中提出的实验旨在深入了解早期Flower 3（ELF 3）的功能，这是一种关键调节器，可调节全天光对生物钟的敏感性，从而使昼夜节律在恒定的光照条件下取得进展。类似的机制也存在于哺乳动物的生物钟中，尽管参与反应的分子成分尚不清楚。这项建议的具体目标是：1.表征ELF 3和已知的生物钟相关蛋白在体内的相互作用。将分析在体外共表达和相互作用的特定时钟蛋白之间的生化和遗传相互作用。2.鉴定与ELF 3相互作用的新蛋白质。酵母双杂交试验和亲和纯化技术与质谱联用将用于鉴定新型ELF 3相互作用蛋白。将通过生物化学和遗传学方法在体内分析鉴定的候选物。相关性：生物钟允许人类日常行为的适当协调，例如睡眠/觉醒周期，以及太阳的昼夜升起和落下。阐明植物生物钟背后复杂分子机制的研究将补充其他系统中的类似分析，最终导致更好地了解人类生物钟，从而治疗昼夜节律紊乱。

DESCRIPTION (provided by applicant): An endogenous molecular oscillator known as the circadian clock controls important biological processes from daily human sleep/wake cycles to plant leaf movements. A major function of the circadian clock is to precisely coordinate physiological responses of an organism to daily environmental changes. The molecular mechanisms underlying the integration of environmental stimuli and the clock are not well understood in many organisms. My objective is to understand how environmental signals are integrated into the circadian clock and the function of specific genes involved in this integration, using Arabidopsis as a model system. Experiments presented in this proposal are designed to provide insight into the function of Early Flowerings (ELF3), a key regulator that regulates the sensitivity of light into the clock throughout the day, allowing for circadian rhythms to progress in constant light conditions. Similar mechanisms exist in the mammalian clock, although the molecular components involved in the response are unknown. The specific aims of this proposal are to: 1. Characterize interactions between ELF3 and known clock associated proteins in vivo. Biochemical and genetic interactions between specific clock proteins that are co-expressed and interact in vitro will be analyzed. 2. Identify novel proteins that interact with ELF3. Yeast two-hybrid assays and affinity purification techniques coupled with mass spectrometry will be used to identify novel ELF3 interacting proteins. Identified candidates will be analyzed by biochemical and genetic approaches in vivo. Relevance: The circadian clock allows for the proper coordination of daily human behaviors, such as sleep/wake cycles, with the diurnal rising and setting of the Sun. Research to elucidate the complex molecular mechanisms underlying the circadian clock in plants will complement similar analyses in other systems, ultimately leading to a better understanding of the circadian clock in humans, allowing for the treatment of circadian disorders.