Investigation of TOC1 function in the Arabidopsis Circadian Clock

拟南芥生物钟中 TOC1 功能的研究

• 批准号: 7874535
• 负责人: Joshua Martin Gendron
• 金额: $ 5.05万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7874535
• 关键词: Affinity; Animal Model; Arabidopsis; Architecture; Attenuated; Behavior; Binding; Biochemical; Biochemistry; Biological Assay; Complex; DNA Binding; Defect; Disease; Eukaryota; Expeditions; Feedback; Genes; Genetic; Genetic Transcription; Genomics; Health; Human; Hybrids; Hypocotyl; Investigation; Libraries; Link; Malignant Neoplasms; Mediating; Modeling; Molecular; Organism; Output; Physiological; Physiology; Platelet Factor 4; Process; Proteins; Regulation; Role; SKP Cullin F-Box Protein Ligases; Set protein; Signal Transduction; System; Techniques; Temperature; Testing; Time; Work; Yeasts; circadian pacemaker; experience; fitness; in vivo; insight; promoter; public health relevance; research study; response; tool; transcription factor; tumor progression; yeast two hybrid system

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.

描述（由申请人提供）：大多数经历日常光/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过程与一天中的特定时间非常重要。重要的是，生物钟在正常运作时可以提供健身优势。生物钟的分子和生化机制尚不清楚，但很明显，生物钟的基本结构在真核生物中是相似的。该提案的目的是利用拟南芥作为模型来研究真核生物钟。具体来说，中央时钟组件 CAB EXPRESSION 1 (TOC1) 的生化作用仍然是拟南芥昼夜节律时钟领域的一个悬而未决的问题。在该提案中，最近发现的 TOC1 相互作用转录因子 CCA1 PROMOTER HIKING EXPEDITION 1 (CHE1) 将用于确定 TOC1 的生化作用。这项工作将连接拟南芥生物钟的核心组件，并将增加我们对真核生物钟架构的理解。具体目标 1. 确定 TOC1 和 ZTL 在 CHE 调节中的作用。 TOC1 和 CHE 的相互作用是 TOC1 和 CCA1 表达控制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 TOC1 和 ZTL 在 CHE 功能调节中的作用。 2.发现调节LHY和CCA1表达的转录复合物。凯实验室创建了拟南芥所有转录因子的文库。该文库将通过酵母双杂交筛选，以寻找与 TOC1 或 ZTL 相互作用的转录因子。将使用一套简化的体内生化和分子技术对与 TOC1/ZTL 相互作用但也调节 LHY 和 CCA1 表达的转录因子进行优先级排序和分类。公共健康相关性：众所周知，生物钟缺陷会导致一系列人类健康相关疾病，生物钟成分参与癌症进展，而癌症会削弱生物钟功能。这项研究的结果将增加我们对真核生物生物钟结构的理解，从而与生物钟相关疾病的研究和治疗相关。