Control of histone methylation during differentiation

分化过程中组蛋白甲基化的控制

• 批准号: 10201923
• 负责人: Michael J Law
• 金额: $ 37.56万
• 依托单位: RICHARD STOCKTON COLLEGE OF NEW JERSEY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201923
• 关键词: Behavior; Biochemical Genetics; Biochemistry; Cell division; Cells; ChIP-seq; Chemicals; Chromatin; Chromatin Remodeling Factor; Complex; Cues; DNA Polymerase II; Data; Defect; Development; Developmental Gene; Enzymes; Epigenetic Process; Equilibrium; Gametogenesis; Gene Expression; Gene Expression Alteration; Gene Expression Profile; Genetic; Genetic Transcription; Genomics; Goals; Histone H3; Histones; Holoenzymes; Individual; Investigation; Laboratories; Lead; Link; Malignant - descriptor; Malignant Neoplasms; Mediating; Mediator of activation protein; Meiosis; Methylation; Methyltransferase; Mitosis; Mitotic; Modeling; Modification; Molecular; Molecular Biology; Molecular Genetics; Monitor; Multienzyme Complexes; Normal Cell; Phase; Phosphorylation; Phosphotransferases; Play; Process; Property; Proteins; Publishing; RNA; Regulation; Role; Saccharomyces cerevisiae; Saccharomycetales; Signal Transduction; Stress; System; Techniques; Testing; Time; Work; Yeasts; cell type; chemical genetics; cyclin C; experimental study; gene product; genetic approach; genome-wide; histone methylation; histone modification; methylation pattern; prevent; programs; recruit; transcription factor; yeast genetics

Differentiation requires cells to integrate external signaling cues with internal cell-type information to execute complex transcriptional programs. Mistakes in this process can lead to developmental defects and cancers. Post-translational histone modifications play central roles in orchestrating all phases of transcription. While much is known regarding how histone modifications are maintained and interpreted, a significant gap exists in our understanding of how the enzyme complexes that catalyze these modifications are controlled. The long-term goal of this project is to dissect the molecular mechanisms that regulate histone modification complexes during differentiation. The specific objective of this proposal is to investigate changes in the integrity, stability, and activity of the histone H3Lys4 methyltransferase COMPASS complex during meiotic differentiation in the budding yeast S. cerevisiae. Our central hypothesis is that there is a two- step mechanism that results in meiosis-specific COMPASS inactivation that is necessary for efficient meiotic entry and completion. The first step implicates changes in locus-specific Set1 methyltransferase recruitment as cells enter meiosis, while the second step involves meiosis- specific Set1 degradation to allow progression past the commitment point. Two Specific Aims are proposed to test this hypothesis. Aim 1 will define the role of locus-specific Set1 antagonism for meiotic entry and execution. Our preliminary and published data indicate that the Cdk8 kinase module of the RNA pol II mediator complex inhibits locus-specific Set1 recruitment. Using molecular, biochemical, and genetic approaches, we will determine if Cdk8-dependent Set1 antagonism occurs via direct or indirect mechanisms. ChIP-sequencing will determine how genome-wide Cdk8-dependent Set1 occupancy is altered as cells enter the meiotic program. Aim 2 will determine how Set1 degradation is incorporated into the meiotic program. Chemical and genetic approaches will decipher the requirement of meiotic gene expression and identify the meiotic hallmarks linked to Set1 degradation. Using genetic and molecular approaches, we will determine the consequences of stabilizing Set1 on meiotic progression and completion while monitoring COMPASS integrity and H3Lys4 me patterns. Successful completion of these Aims will have a significant impact as they will provide mechanistic detail into how COMPASS is retooled during differentiation. The strength of this project is that it merges classical techniques in yeast genetics, biochemistry, and molecular biology with contemporary approaches in genomics and computation.

分化需要细胞将外部信号信号与内部细胞类型信息相结合 来执行复杂的转录程序。这一过程中的错误可能会导致 缺陷和癌症。翻译后的组蛋白修饰在编排中起着核心作用 转录的所有阶段。虽然关于组蛋白修饰是如何 维持和解释，我们对酶的理解存在着显著的差距 催化这些修饰的络合物是受控制的。这个项目的长期目标是 解析组蛋白修饰复合体调控的分子机制 差异化。这项提案的具体目标是调查诚信方面的变化， 组蛋白H3Lys4甲基转移酶COMPASS复合体在减数分裂过程中的稳定性和活性 芽生酵母菌的分化。我们的中心假设是有两个- 导致特定于减数分裂的COMPASS失活的阶跃机制，这是 有效地进入和完成减数分裂。第一步意味着特定于基因座的Set1的改变 当细胞进入减数分裂时，甲基转移酶重新招募，而第二步涉及减数分裂- 特定的SET 1降级，以允许进展超过承诺点。两个具体目标是 提出用来检验这一假说。目标1将定义位点特异的Set1拮抗作用 减数分裂进入和执行。我们的初步和已发表的数据表明，CDK8激酶 RNA PolII介体复合体的模块抑制位点特异性的Set1招募。vbl.使用 分子、生化和遗传方法，我们将确定CDK8依赖的Set1 对抗通过直接或间接机制发生。芯片测序将决定如何 全基因组依赖CDK8的Set1的占有率随着细胞进入减数分裂程序而改变。目标 2将决定Set1降解如何并入减数分裂计划。化学药品和 遗传学方法将破译减数分裂基因表达的要求，并识别 与Set1降解有关的减数分裂特征。使用遗传和分子方法，我们将 确定稳定Set1对减数分裂进程和完成的影响，同时 监控指南针的完整性和H3Lys4 Me模式。圆满完成这些目标 将产生重大影响，因为它们将提供有关指南针如何的机械细节 在差异化期间进行了重新装备。这个项目的优点在于它融合了经典的技术 酵母遗传学、生物化学和分子生物学与当代基因组学方法 和计算。