Investigate the function of NDF in chromatin dynamics and gene expression

研究 NDF 在染色质动力学和基因表达中的功能

基本信息

批准号：
10587031
负责人：
Jia Fei
金额：
$ 32.97万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-15 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10587031
关键词：
Address Affinity Chromatography Area Biochemical Biochemistry Bioinformatics Biological Assay Biological Phenomena CRISPR/Cas technology Cell Nucleus Cell Survival Cell physiology Cells Chromatin Collaborations Cryoelectron Microscopy DNA Development Disease Environment Gene Expression Genes Genetic Transcription Genomics Goals Health Histones Homologous Gene Human In Vitro Knock-out Knowledge Lethal Genes Life Mediating Methods Molecular Nucleosomes Outcome Pathway interactions Play Polymerase Positioning Attribute Process Proteins RNA Polymerase II Regulation Role Somatic Cell Structure Techniques Testing Transcription Elongation Work Yeasts biophysical techniques cell fate specification genome-wide genome-wide analysis insight laser tweezer novel optic tweezer pancreatic differentiation 2 protein recruit single molecule structural biology transcription factor ultra high resolution

项目摘要

Project Summary/Abstract Our current understanding of chromatin transcription has been limited by our knowledge of the factors that are involved in this process. In this regard, we have identified a novel nucleosome destabilization factor, termed NDF, that can destabilize nucleosomes and facilitate transcription elongation through nucleosomes. In this proposal, we will investigate the function of NDF in chromatin dynamics and RNA Polymerase II transcription elongation using biochemistry, genomics, bioinformatics and structural biology approaches. In Specific Aim 1, we will develop and employ new methods and strategies to study the molecular mechanism of NDF-mediated nucleosome destabilization and RNA Polymerase II transcription. We will use ultra-high-resolution optical tweezer as well as cryo-electron microscopy to characterize the dynamics of NDF-mediated nucleosome destabilization. In Specific Aim 2, we will examine how NDF is recruited to the transcribed regions of active genes in cells. We will assess whether other cellular factors, other than H3K36me3 histones, contribute to the recruitment of NDF. In Specific Aim 3, we will determine how genes lacking NDF are transcribed, and investigate protein factors that are related to NDF. The studies proposed here should significantly increase our understanding of the mechanisms of Pol II transcription elongation through chromatin.

项目概要/摘要我们目前对染色质转录的理解受到我们对染色质转录因素的了解的限制。参与这个过程。在这方面，我们发现了一种新的核小体不稳定因子，称为 NDF，可以破坏核小体的稳定性并通过核小体促进转录延伸。在这个提案中，我们将研究 NDF 在染色质动力学和 RNA 聚合酶 II 转录中的功能使用生物化学、基因组学、生物信息学和结构生物学方法进行延伸。在具体目标 1 中，我们将开发和采用新的方法和策略来研究NDF介导的分子机制核小体不稳定和 RNA 聚合酶 II 转录。我们将使用超高分辨率光学镊子以及冷冻电子显微镜来表征 NDF 介导的核小体的动力学不稳定。在具体目标 2 中，我们将研究 NDF 如何被招募到活性转录区域细胞中的基因。我们将评估除 H3K36me3 组蛋白之外的其他细胞因子是否有助于 NDF 的招募。在具体目标 3 中，我们将确定缺乏 NDF 的基因如何转录，以及研究与 NDF 相关的蛋白质因素。这里提出的研究应该会显着提高我们的了解通过染色质进行 Pol II 转录延伸的机制。