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

• 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
• 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

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.

项目总结/摘要 我们目前对染色质转录的理解受到我们对影响转录的因素的认识的限制。 参与了这个过程。在这方面，我们已经确定了一种新的核小体不稳定因子，称为 NDF，其可以使核小体不稳定并促进通过核小体的转录延伸。在这 本研究拟探讨NDF在染色质动力学和RNA聚合酶II转录中的作用 延伸使用生物化学、基因组学、生物信息学和结构生物学方法。具体目标1、 我们将开发和采用新的方法和策略来研究NDF介导的 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是如何被募集到活性转录区的。 细胞中的基因我们将评估除了H3 K36 me 3组蛋白以外的其他细胞因子是否有助于H3 K36 me 3的表达。 招募NDF。在具体目标3中，我们将确定缺乏NDF的基因如何转录， 研究与NDF相关的蛋白质因子。这里提出的研究应该大大增加我们的 了解Pol II转录通过染色质延伸的机制。