Identification and Analysis of Circadian Clock-Controlled Genes

生物钟控制基因的鉴定和分析

基本信息

批准号：
8788365
负责人：
JENNIFER J. LOROS
金额：
$ 71.6万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
1989
资助国家：
美国
起止时间：
1989-09-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8788365
关键词：
Adipocytes Adipose tissue Aspergillosis Aspergillus fumigatus Biochemical Genetics Biochemistry Bioinformatics Biological Biological Assay Biological Clocks Biological Models Biology Cell Line Cell physiology Cells ChIP-seq Circadian Rhythms Clock protein Diabetes Mellitus Disease Environment Epitopes Eukaryota Eukaryotic Cell Feedback Foundations Gene Expression Gene Expression Profile Genes Genetic Goals Grant Human Human body Informatics Kinetics Knock-out Language Life Light Mammalian Cell Maps Mental disorders Metabolic Metabolic syndrome Metabolism Molecular Genetics Motion Mouse Cell Line Mus Mycoses NRIP1 gene Neurospora Nuclear Receptors Organism Output Pathway interactions Patients Photobiology Photoreceptors Physiology Process Proteins Protocols documentation RNA Sequences Reagent Regulation Reporter Role Structure System Techniques Time Transcriptional Regulation Work cell behavior cell type chromatin immunoprecipitation circadian pacemaker empowered fungus human NRIP1 protein insight lipid metabolism next generation sequencing pathogen response skills tool transcription factor transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): Our long term goal is to describe in the language of genetics and biochemistry the feedback cycles and pathways that comprise intracellular circadian systems - how they work, how they are synchronized with the environment, and how time information generated by them is used to regulate the behavior of cells. This proposal focuses on the model system Neurospora, as well as on the mouse and mammalian cell lines, to understand the paradigms underlying circadian control of cell physiology and metabolism. We also continue a longstanding effort aimed at understanding circadian photoreception and photobiology in Neurospora and use this to break new ground on a salient fungal pathogen. In Specific Aim 1, we will carry out a global analysis and description of the circadian output network in Neurospora, using RNA sequencing, chromatin immunoprecipitation, and bioinformatics to describe the regulatory hierarchy governing circadian regulation of transcription in a cell. Then, using the exceptional background of biochemical genetics available in Neurospora, we will track the metabolic activities carried out by the proteins encoded by clock-controlled genes, and as foci of clock-controlled activities emerge, we will begin to lay the spectrum of clock-controlled processes on the Neurospora metabolic map to see how the clock regulates metabolism and physiology. In Specific Aim 2, we will exploit recent atomic level structure analysis of the photoreceptive domain to determine the biological significance of photocycle kinetics. We will extend analysis of photobiology to the important fungal pathogen Aspergillus fumigatus where we can envision a way to exploit our understanding of fungal photobiology to enable a new treatment for hundreds of thousands of patients with aspergillosis. In Specific Aim 3, we will use RNA sequencing to determine the circadian profile of clock-controlled genes in adipocytes of wt and RIP140 knockout cells, and will use chromatin immunoprecipitation of RIP140 in adipocytes to begin to dissect the role of this co-activator/co-repressor in the circadian biology of this important cell type. We will look for physical association of RIP140 with known clock proteins and transcription factors involved with circadian output, and will generate white adipose tissue-specific knockouts of the clock to probe the significance of circadian regulation to fat metabolism in the mouse, hoping to gain insights into diabetes and metabolic syndrome. These projects are complementary and mutually enriching in that they each rely on genetic and molecular techniques to dissect, and ultimately to understand, the response of cells to their environment and the organization of eukaryotic cells as a function of time.

描述（由申请人提供）：我们的长期目标是用遗传学和生物化学的语言描述包含细胞内昼夜节律系统的反馈周期和途径 - 它们的工作方式，如何与环境同步，以及如何使用它们的时间信息来调节细胞的行为。该建议着重于模型系统神经孢子以及小鼠和哺乳动物细胞系，以了解细胞生理学和代谢的昼夜节律控制的范例。我们还继续进行长期的努力，旨在理解神经孢菌中的昼夜节律光感受和光生物学，并利用它在显着的真菌病原体上打破了新的地面。在特定目标1中，我们将使用RNA测序，染色质免疫沉淀和生物信息学对神经孢子中的昼夜节律输出网络进行全球分析和描述，以描述细胞中昼夜节律转录调节的调节层次结构。 Then, using the exceptional background of biochemical genetics available in Neurospora, we will track the metabolic activities carried out by the proteins encoded by clock-controlled genes, and as foci of clock-controlled activities emerge, we will begin to lay the spectrum of clock-controlled processes on the Neurospora metabolic map to see how the clock regulates metabolism and physiology. 在特定目标2中，我们将利用光感受体结构域的最新原子水平结构分析，以确定光循环动力学的生物学意义。我们将将光生物学的分析扩展到重要的真菌病原体曲霉菌，在那里我们可以设想一种方法来利用我们对真菌光生物学的理解，从而为成千上万的曲霉病患者提供新的治疗方法。在特定的目标3中，我们将使用RNA测序来确定WT和RIP140基因敲除细胞中时钟控制基因的昼夜节律谱，并将使用脂肪细胞中RIP140的染色质免疫沉淀来开始与该cile-Repress in CircadAdian in CircadAdAdian in CircadAdAdian in CircadAdAdian in CircadAdAdAdAdAdy in CircadAdAdAdAdy in CircadAdAdAdy in CircadAdAdAdian in CircadAdAdAdy cignadAdAdAdy的作用。我们将寻找RIP140与已知的时钟蛋白和与昼夜节律输出有关的转录因子的物理关联，并将产生白色脂肪组织特异性敲除时钟的特异性敲除，以探测昼夜小鼠调节对小鼠脂肪代谢的重要性，希望能够对糖尿病和代谢综合征的洞察力。这些项目是互补的和相互丰富的，因为它们每个人都依靠遗传和分子技术来剖析细胞对环境的反应以及真核细胞随时间的影响。