Investigation of TOC1 function in the Arabidopsis Circadian Clock

描述（由申请人提供）：大多数经历每日光/暗或温度循环的生物体都有一个称为生物钟的分子振荡器。生物钟对于协调日常生理活动和一天中特定时间的过程很重要。重要的是，当生物钟正常工作时，它提供了健身优势。生物钟的分子和生物化学机制还没有很好的理解，但很明显，生物钟的基本结构在真核生物中是相似的。本研究的目的是以拟南芥为模型研究真核生物的生物钟。具体而言，生物化学作用的中央时钟组件，定时CAB表达1（TOC 1），仍然是一个悬而未决的问题，在拟南芥昼夜节律时钟领域。在这项提议中，最近发现的TOC 1相互作用的转录因子，CCA 1启动子提升扩展1（CHE 1）将被用来确定TOC 1的生化作用。这项工作将连接拟南芥生物钟的核心组成部分，并将增加我们对真核生物钟结构的理解。具体目标1.确定TOC 1和ZTL在CHE调节中的作用。TOC 1和CHE的相互作用是TOC 1和控制CCA 1表达之间的第一个生化联系。生物化学，瞬时转录测定和遗传学将用于确定TOC 1和ZTL在CHE功能调节中的作用。2.发现调节LHY和CCA 1表达的转录复合物。Kay实验室已经建立了一个包含拟南芥所有转录因子的文库。利用酵母双杂交技术筛选与TOC 1或ZTL相互作用的转录因子。一套精简的体内生物化学和分子技术将被用来优先考虑和分类的转录因子与TOC 1/ZTL相互作用，但也调节LHY和CCA 1的表达。公共卫生相关性：已知生物钟缺陷会导致一系列人类健康相关疾病，并且生物钟成分参与癌症进展，并且癌症减弱生物钟功能。本研究的结果将增加我们对真核生物中生物钟结构的理解，从而将与生物钟相关疾病的研究和治疗相关。

DESCRIPTION (provided by applicant): Most organisms that experience daily light/dark or temperature cycles have a molecular oscillator called the circadian clock. The circadian clock is important for coordinating daily physiological activities and processes with specific times of day. Importantly, the circadian clock provides fitness advantage when it functions correctly. The molecular and biochemical mechanisms of the circadian clock are not well understood, but it is clear that the basic architecture of circadian clocks is similar in eukaryotes. The objective of this proposal is to use Arabidopsis as a model to study eukaryotic circadian clocks. Specifically, the biochemical role of the central clock component, TIMING OF CAB EXPRESSION 1 (TOC1), remains an outstanding question in the Arabidopsis circadian clock field. In this proposal, the recently discovered TOC1-interacting transcription factor, CCA1 PROMOTER HIKING EXPEDITION 1 (CHE1) will be used to determine the biochemical role of TOC1. This work will connect the core components of the Arabidopsis circadian clock and will increase our understanding of eukaryotic clock architecture. Specific Aims 1. Determine the roles of TOC1 and ZTL in CHE regulation. The interaction of TOC1 and CHE is the first biochemical link between TOC1 and control of CCA1 expression. Biochemistry, transient transcription assays, and genetics will be used to determine the role of TOC1 and ZTL in regulation of CHE function. 2. Discovery of transcriptional complexes that regulate LHY and CCA1 expression. A library of all transcription factors from Arabidopsis has been created in the Kay lab. This library will be screened by yeast two-hybrid to find transcription factors that interact with TOC1 or ZTL. A streamlined set of in vivo biochemical and molecular techniques will be used to prioritize and categorize the transcription factors that interact with TOC1/ZTL but also regulate LHY and CCA1 expression. Public Health Relevance: Circadian clock defects are known to result in a range of human health related disorders, and clock components are involved in cancer progression and cancer attenuates clock function. The results of this study will increase our understanding of circadian clock architecture in eukaryotes and thus will be relevant to the study and treatment of clock related diseases.