Dynamics of epigenetic marks on newly replicated chromatin

新复制的染色质上表观遗传标记的动态

• 批准号: 7740766
• 负责人: Stephen J. Kron
• 金额: $ 19.5万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-20 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7740766
• 关键词: Adopted; Affinity Chromatography; Aging; Alkynes; Alleles; Azides; Binding; Binding Proteins; Binding Sites; Biological Assay; Biotin; Bromodeoxyuridine; Cataloging; Catalogs; Cell Cycle; Cell Cycle Arrest; Cell Cycle Progression; Cell division; Cells; Chemicals; Chemistry; Chromatin; Chromosomes; Clone Cells; Complex; DNA; DNA Methylation; DNA Repair; DNA Sequence; DNA biosynthesis; DNA-Directed DNA Polymerase; Data; Deoxyuridine; Deposition; Development; Diacetyl; Disease; Epigenetic Process; Event; Fluorescent Probes; Gene Expression; Gene Silencing; Generations; Genes; Genetic Transcription; Genomics; Growth; Hereditary Disease; Heterochromatin; Histone Acetylation; Histone H4; Histones; Imaging Device; Individual; Informatics; Interphase; Label; Learning; Link; Maintenance; Malignant Neoplasms; Mediator of activation protein; Methods; Methylation; Mitosis; Mitotic; Modification; Molecular; Molecular Chaperones; Mutation; Normal Cell; Nucleosomes; Pattern; Phase; Phenotype; Physical condensation; Physiologic pulse; Polymerase; Positioning Attribute; Post-Translational Protein Processing; Proteins; Proteolysis; Proteomics; RNA Interference; Reaction; Recycling; Regulator Genes; Resolution; Role; Site; Stretching; Testing; Therapeutic; Time; Tissues; Trypsin; Variant; base; cancer therapy; cell type; chromatin modification; chromatin protein; crosslink; cycloaddition; histone modification; imprint; inhibitor/antagonist; insight; interest; mass spectrometer; mutant; novel; public health relevance; regenerative therapy; repaired; restoration; senescence; small molecule; success; theories; tool; transcription factor; tumor

DESCRIPTION (provided by applicant): Although the effort to catalog the full range of epigenetic marks and assign them functional roles has been a remarkable success to date, how epigenetic states persist through replication and mitosis (epigenetic inheritance) remain as important open questions. Current theories suggest that marked histones remain associated with DNA during chromosomal replication and mitotic condensation and then serve as templates for reassembly of the interphase chromatin state. In the case of DNA replication, the requirement for DNA strands to be unwound and accessed by polymerases places severe constraints on potential mechanisms for persistence of epigenetic marks. Recent data have implicated histone chaperones in concert with chromatin remodelers and DNA polymerase complexes, as the mediators of nucleosome dissasembly and reassembly at sites of replication. However, much needs to be learned about how recycled and/or newly synthesized histones are deposited and remodeled to adopt the correct patterns of histone types and modifications. We are interested in dissecting the inheritance of epigenetic marks by exploiting proteomic approaches to analyze maturation and dynamics of replicated chromatin during S phase and mitosis. As such, we will develop methods to obtain marked chromatin via pulse chase analysis to isolate sites of DNA synthesis or chromatin at different intervals after replication. We will particularly focus on the inheritance of heterochromatin. Then, we will use quantitative proteomic analysis to follow the time-course of changes in chromatin proteins and modifications following replication, through mitosis and into the next cell cycle. Finally, we hope to determine which proteins and modifications are required for inheritance of chromatin states through replication and persistence of chromatin marks after mitosis. PUBLIC HEALTH RELEVANCE: We propose to better understand epigenetic inheritance by documenting the molecular events that allow newly replicated or repaired chromatin to return to its former state. Discovery of the specific mechanisms that allow patterns of expression to be passed on during normal cell division will have broad applications in understanding and treating diseases of development and aging and may identify new targets for cancer therapy.

描述（由申请人提供）：尽管对全部表观遗传标记并分配功能角色的努力一直是迄今为止取得的非凡成功，但表观遗传状态如何通过复制和有丝分裂（表观遗传遗传遗传）仍然是重要的开放问题。当前的理论表明，在染色体复制和有丝分裂凝结过程中，标记的组蛋白仍然与DNA相关，然后用作重新组装相间染色质态的模板。在DNA复制的情况下，DNA链被解开并被聚合酶访问的要求对表观遗传标记的持续存在严重限制。最近的数据已与染色质重塑剂和DNA聚合酶复合物共同表示组蛋白伴侣，是核小体隔离的介体在复制部位的介体和重新组装。但是，需要了解如何将回收和/或新合成的组蛋白沉积和重塑以采用组蛋白类型和修饰的正确模式。我们有兴趣通过利用蛋白质组学方法来分析S相和有丝分裂的蛋白质组学方法来解剖表观遗传标记的遗传。因此，我们将开发通过脉冲追逐分析获得明显的染色质的方法，以在复制后以不同的间隔分离DNA合成或染色质的位点。我们将特别关注异染色质的遗传。然后，我们将使用定量蛋白质组学分析遵循染色质蛋白变化的时间顺序，并通过有丝分裂和下一个细胞周期进行复制后修饰。最后，我们希望通过有丝分裂后的复制和持久性来确定染色质状态遗传需要哪些蛋白质和修饰。公共卫生相关性：我们建议通过记录允许新复制或修复的染色质返回其以前州的分子事件来更好地理解表观遗传的遗传。发现允许在正常细胞分裂期间传递表达模式的特定机制将在理解和治疗发育和衰老疾病中广泛应用，并可能确定癌症治疗的新靶标。