Molecular analysis of circadian regulation in Arabidopsis

拟南芥昼夜节律调节的分子分析

基本信息

批准号：
8706893
负责人：
Stacey L. Harmer
金额：
$ 28.6万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-03-01 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8706893
关键词：
Affect Affinity Animal Model Arabidopsis Architecture Behavior Binding Biochemical Biochemical Genetics Biological Clocks Biological Models Biological Process Chromatin Circadian Rhythms Clock protein Complex Development Disease Elements Ensure Environment Equilibrium Eukaryota Eukaryotic Cell Family Feedback Fission Yeast Gene Expression Gene Expression Profile Gene Expression Regulation Gene Targeting Genes Genetic Genetic Models Genome Genomics Goals Health Homeostasis Homologous Gene Hour Human Individual Light Malignant Neoplasms Mammals Modeling Molecular Molecular Analysis Mood Disorders Mouse-ear Cress Mutation Nature Organism Output Pathway interactions Pattern Phase Physiological Processes Physiology Plant Components Plant Model Play Process Proteins Protocols documentation Regulation Regulatory Pathway Resources Role Shapes Sleep Disorders Social Welfare System Taxon Techniques Testing Time Transcription Coactivator Transcriptional Regulation Work base biochemical model chromatin immunoprecipitation circadian pacemaker fitness genetic regulatory protein genome-wide in vivo insight mathematical model member mutant novel promoter public health relevance research study response transcription factor

项目摘要

DESCRIPTION (provided by applicant): Circadian rhythms provide organisms with an adaptive advantage, enhancing the health and fitness of individuals by ensuring that diverse physiological processes occur at the most appropriate times of day. The long-term objective of this proposal is to understand the molecular basis of circadian rhythms in eukaryotic cells. We have identified key regulators of the circadian clock in our previous studies, and will now use genomic, biochemical, genetic, and mathematical modeling approaches to appropriately place these proteins in the circadian system. We will conduct our studies in Arabidopsis thaliana, a model plant that is uniquely well-suited for these experiments due to its compact genome, extensive genetic and genomic resources, and ability to tolerate mutations in chromatin regulatory pathways that are lethal to other complex eukaryotes. We will first use genomic, biochemical, and genetic approaches to characterize the role of a protein conserved across eukaryotes (but of unknown biochemical function) in the regulation of chromatin. We anticipate this work will generate insights into mechanisms governing chromatin organization and thus gene expression in diverse eukaryotes. Next, we will use genetic and biochemical techniques to investigate the roles of a family of related transcription factors in the workings of the circadian oscillator. In the process, we will test predictions made by a mathematical model, evaluating how well this model describes the regulatory relationships that drive the clock. Finally, we will modify an existing competitive chromatin immunoprecipitation protocol to investigate how the binding dynamics of antagonistic transcription factors to chromatin shape the dynamic regulation of gene expression in vivo. These experiments will reveal general principles governing the temporal regulation of gene expression and thus will be applicable to many organisms and processes. Our exploration of shared mechanisms that control circadian clock function in diverse organisms will increase our understanding of how clocks function in humans and shed light on how they promote human health and welfare.

描述（由申请人提供）：昼夜节律为有机体提供适应性优势，通过确保在一天中最合适的时间发生多种生理过程，从而增强了个体的健康和适应性。该提议的长期目标是了解真核细胞中昼夜节律的分子基础。我们已经在以前的研究中确定了昼夜节律时钟的关键调节剂，现在将使用基因组，生化，遗传和数学建模方法来适当地将这些蛋白质置于昼夜节律系统中。我们将在拟南芥（Arabidopsis thaliana）进行研究，这是一种模型植物，由于其紧凑的基因组，广泛的遗传和基因组资源，以及能够在染色质调节途径中耐受致死的染色质调节途径中的突变的能力，因此非常适合这些实验。我们将首先使用基因组，生化和遗传方法来表征跨真核生物（但生化功能未知的）在染色质调节中保守的作用（但生化功能未知）。我们预计，这项工作将产生有关染色质组织的机制，从而在各种真核生物中的基因表达。接下来，我们将使用遗传和生化技术来研究相关转录因子家族在昼夜节律中的作用振荡器。在此过程中，我们将测试通过数学模型做出的预测，以评估该模型描述驱动时钟的调节关系的程度。最后，我们将修改现有的竞争性染色质免疫沉淀方案，以研究拮抗转录因子对染色质的结合动力学如何塑造体内基因表达的动态调节。这些实验将揭示有关基因表达时间调节的一般原则，因此将适用于许多生物体和过程。我们对控制昼夜节律在各种生物中功能的共同机制的探索将增加我们对时钟在人类中的功能的理解，并阐明它们如何促进人类健康和福利。