Examining Circadian Changes in Genome Structure

检查基因组结构的昼夜变化

• 批准号: 8459989
• 负责人: WILLIAM J BELDEN
• 金额: $ 27.4万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8459989
• 关键词: Animal Model; Biochemistry; Biological Clocks; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromatin Structure; Circadian Rhythms; Cis-Acting Sequence; Complex; DNA; DNA Methylation; DNA Sequence; Data; Defect; Ensure; Enzymes; Feedback; Frequencies; Functional RNA; Gene Expression; Gene Frequency; Genes; Genetic; Genetic Transcription; Genome; Global Change; Health; Heterochromatin; Histone H3; Histones; Human; Indium; Lysine; Mass Spectrum Analysis; Metabolism; Methylation; Methyltransferase; Modification; Molecular; Neurospora; Normal Cell; Nucleosomes; Organism; Output; Pathway interactions; Phase; Play; Positioning Attribute; Process; Proteins; Regulation; Research; Role; Running; Structure; System; Technology; Time; Transcript; Transcription Coactivator; chromatin modification; chromatin remodeling; circadian pacemaker; design; genome-wide; histone modification; innovation; insight; interest; method development; mutant; programs; promoter; research study

DESCRIPTION (provided by applicant): Research over the past decade has revealed an important role for chromatin-modifying and chromatin-remodeling enzymes in coordinated regulation of the circadian clock. This is not unexpected given that a core element in eukaryotic circadian clocks is a transcriptional negative feedback loop. Chromatin functions to organize eukaryotic DNA, but also creates a barrier for transcription. This necessitates a role for chromatin remodeling in maintaining circadian rhythms. In this proposal, I am seeking to further define the modifications that occur at the Neurospora central clock gene frequency and examine the role of heterochromatin in clock and clock-controlled gene expression. Experiments discussed herein will examine this problem using a combined approach. In Specific Aim 1, I propose to isolate the central clock gene frequency in its native chromatin state and identify chromatin modification by Mass Spectrometry. In Specific Aim 2, I will examine how the KMT1 DIM-5 is directed to chromatin and the role of H3K9 methylation in clock and clock controlled gene expression. In Specific Aim 3, I examine if there are circadian clock-regulated changes to genome structure. Data obtained in these aims will be compared and contrasted to understand the relationship between genome structure and clock regulated gene expression. Overall, this proposal will further our understanding of how chromatin structural changes effects circadian regulated gene expression.

描述（由申请人提供）：过去十年的研究揭示了染色质修饰和染色质重塑酶在协调调节生物钟中的重要作用。这并不意外，因为真核生物生物钟的核心元素是转录负反馈环。染色质的功能是组织真核生物的DNA，但也为转录创造了障碍。这需要染色质重塑在维持昼夜节律中的作用。在这个提议中，我试图进一步定义发生在脉孢菌中心时钟基因频率的修饰，并研究异染色质在时钟和时钟控制的基因表达中的作用。本文讨论的实验将使用组合方法来检查这个问题。在具体目标1中，我建议在其天然染色质状态下分离中心时钟基因频率，并通过质谱法鉴定染色质修饰。在具体目标2中，我将研究KMT 1 DIM-5是如何定向到染色质的，以及H3 K9甲基化在时钟和时钟控制的基因表达中的作用。在具体目标3中，我检查了基因组结构是否存在生物钟调节的变化。在这些目标中获得的数据将进行比较和对比，以了解基因组结构和时钟调控基因表达之间的关系。总的来说，这一建议将进一步我们了解染色质结构的变化如何影响昼夜节律调控的基因表达。