The Mute button: Turning down the volume of histone expression

静音按钮：调低组蛋白表达的音量

• 批准号: 10750147
• 负责人: Mark Stephen Geisler
• 金额: $ 4.77万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750147
• 关键词: ATAC-seq; Affect; Anabolism; Binding; Biological Models; Biotin; C-terminal; Cell Cycle; Cell Cycle Arrest; Cell Cycle Progression; Cell Proliferation; Cell physiology; Cells; Cellular biology; Chromatin; Chromatin Structure; Code; Coupled; Couples; DNA; DNA Damage; DNA Packaging; DNA biosynthesis; Data; Development; Diploidy; Drosophila Proteins; Drosophila genus; Embryo; Eukaryota; Eukaryotic Cell; G2 Phase; Gene Cluster; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Genomics; Goals; Histones; Homologous Gene; Human; Knowledge; Label; Lead; Liquid substance; M cell; Mass Spectrum Analysis; Mediating; Messenger RNA; Molecular; Mutation; Normal Cell; Nucleosomes; Organism; Phase; Phenotype; Point Mutation; Postembryonic; Process; Production; Proliferating; Protein Biosynthesis; Proteins; Proteomics; Radial; Regulation; Repression; Research; Role; S phase; SANT Domain; Structure; Techniques; Testing; Tissues; Toxic effect; Transcript; Transcriptional Activation; Variant; Work; Yeasts; cell type; design; embryo tissue; experimental study; fluorescence imaging; follow-up; gene repression; genetic manipulation; genome integrity; insight; interdisciplinary approach; mosaic; mutant; null mutation; tool

PROJECT SUMMARY During DNA replication, a cell must replicate not only its DNA, but also the nucleosomes that package its genome. To meet this extremely high demand for rapid histone protein biosynthesis, all eukaryotes coordinate high expression of replication-dependent (RD) histone gene clusters during S-phase of the cell division cycle. Regulating the level of expression of RD-histone genes during S-phase is important for maintaining genomic integrity and normal cell cycle progression, as too many or too few histones lead to toxic effects such as enhanced DNA damage sensitivity or cell cycle arrest. While the transcriptional activation of RD-histone genes has been characterized, little is known about the negative regulation that occurs as cells exit S-phase or the modulation of transcription that occurs during S-phase. My preliminary research has shown the Drosophila protein Mute (a homologue of human Yarp/Gon4L) is a prime candidate for a negative transcriptional regulator of RD-histone genes. I have found that loss of Mute uncouples expression of RD-histone genes from S-phase in Drosophila embryos. I hypothesize Mute is both restricting RD-histone expression to S-phase as well as regulating levels of expression during S- phase. This proposal seeks to uncover the mechanisms through which Mute represses RD- histone genes and how this repression is connected to the cell cycle. Using an interdisciplinary approach, the mechanisms of Mute’s repression at both the cellular and molecular levels will be explored using a combination of genomic, fluorescent imaging, genetic, and proteomic techniques in the Drosophila model system. This project will enhance our understanding of the role of histone gene regulation in metazoans and provide insight into the mechanisms that govern the regulation of this highly conserved cellular process.

项目摘要 在DNA复制过程中，细胞不仅要复制其DNA，还要复制 包装它的基因组。为了满足这种对快速组蛋白生物合成的极高需求， 所有真核生物都协调复制依赖性（RD）组蛋白基因簇的高表达 在细胞分裂周期的S期。调节RD-组蛋白基因的表达水平 在S期期间对于维持基因组完整性和正常细胞周期进程是重要的， 因为太多或太少的组蛋白会导致毒性效应，例如增强的DNA损伤敏感性 或细胞周期停滞。虽然RD-组蛋白基因的转录激活已经被证实是一种重要的基因表达调控机制， 特征，很少有人知道的负调控发生细胞退出S期或 在S期发生的转录调节。我的初步研究表明 果蝇蛋白Mute（人Yarp/Gon 4L同源物）是一种主要候选蛋白， RD-组蛋白基因的负转录调节因子。我发现失去哑巴 果蝇胚胎S期RD组蛋白基因的表达。我假设哑巴两者都是 将RD-组蛋白表达限制在S期以及调节S期表达水平， 相位该提案旨在揭示Mute抑制RD的机制- 组蛋白基因以及这种抑制是如何与细胞周期联系在一起的。使用跨学科的 方法，在细胞和分子水平上Mute的抑制机制将被 利用基因组学、荧光成像、遗传学和蛋白质组学技术的组合进行探索 在果蝇模型系统中。这个项目将增强我们对组蛋白作用的理解 多细胞动物中的基因调控并提供对调控机制的深入了解 这一高度保守的细胞过程。