Identification and Analysis of Circadian Clock-Controlled Genes

生物钟控制基因的鉴定和分析

• 批准号: 9251857
• 负责人: JENNIFER J. LOROS
• 金额: $ 63.01万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9251857
• 关键词: Adipocytes; Alpha Cell; Aspergillosis; Aspergillus fumigatus; Biochemical Genetics; Biochemistry; Bioinformatics; Biological Clocks; Biological Models; Biology; Cell Line; Cell physiology; Cells; Cellular Metabolic Process; Circadian Rhythms; Diabetes Mellitus; Disease; Environment; Eukaryotic Cell; Feedback; Genes; Genetic; Goals; Human; Human body; Incidence; Jet Lag Syndrome; Knockout Mice; Knowledge; Language; Light; Mammalian Cell; Mammals; Maps; Mental disorders; Metabolic; Metabolic syndrome; Metabolism; Molecular; Mus; NRIP1 gene; Neurospora; Output; Pathway interactions; Patients; Photobiology; Photoreceptors; Physiology; Process; Proteins; Regulation; Role; System; Techniques; Testing; Time; Transcriptional Regulation; Work; cell behavior; cell type; chromatin immunoprecipitation; circadian pacemaker; experimental study; fungus; immune function; insight; lipid metabolism; macrophage; mutant; pathogen; public health relevance; response; transcriptome; transcriptome sequencing

 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. Our work has three foci. One Focus anticipates 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 the second Focus, we will apply our knowledge of circadian output pathways to mammalian cells, using RNA sequencing to determine the circadian profile of clock-controlled genes in adipocytes and in macrophages from wt and RIP140 knockout mice. We will use RNA-seq to characterize the transcriptomes of WT and mutant cells and use chromatin immunoprecipitation to begin to dissect the role of this co-activator/co-repressor in the circadian biology of these important cell types. These experiments will probe the significance of circadian regulation to fat metabolism and to immune function in a mammal, with the hope of gaining insights into the incidence in humans of diabetes, metabolic syndrome, time-of-day differences in immune function. A third Focus is in photobiology. We will explore the mechanism whereby light activates the principal fungal photoreceptor, and extend analysis of photobiology in the important fungal pathogen Aspergillus fumigatus. We envision and will test a way to exploit our understanding of fungal photobiology to enable a new treatment for hundreds of thousands of patients with aspergillosis. 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（由适用提供）：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。我们的工作有三个焦点。一个重点预期使用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。然后，使用Neurospora中可用的生化遗传学的特殊背景，我们将跟踪由时钟控制基因编码的蛋白质进行的代谢活性，并且作为时钟控制活动的焦点，我们将开始放置。 神经孢子代谢图上时钟控制过程的频谱，以查看时钟如何调节代谢和生理。在第二个焦点中，我们将使用RNA测序将昼夜节律输出途径的知识应用于哺乳动物细胞，以确定脂肪细胞中时钟控制基因的昼夜节律分布以及WT和RIP140敲除小鼠的巨噬细胞中的昼夜节目。我们将使用RNA-Seq表征WT和突变细胞的转录组，并使用Chormetin免疫沉淀开始剖析该共激活剂/共抑制器在这些重要细胞类型的昼夜节律生物学中的作用。这些实验将探究昼夜节律对脂肪代谢和免疫功能的重要性，希望能够洞悉糖尿病人类，代谢综合征，免疫功能的时间差异。第三个重点是光生物学。我们将探索光激活主要真菌感光器的机制，并扩展重要真菌病原体曲霉曲霉中光生物学的分析。我们设想并将测试一种利用我们对真菌光生物学的理解的方法，以便为成千上万的曲霉病患者提供新的治疗方法。这些项目是完整的和相互富裕的，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着时间的影响。