Histone Modifications During Embryogenesis

胚胎发生过程中的组蛋白修饰

• 批准号: 8320462
• 负责人: ALEXANDER F SCHIER
• 金额: $ 24.92万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-15 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8320462
• 关键词: Acetylation; Address; Animals; Attention; Biological Assay; Biology; Biomedical Research; Cells; Chimeric Proteins; Chromatin; Chromatin Structure; Congenital Abnormality; DNA; Degenerative Disorder; Development; Developmental Gene; Disease; Embryo; Embryonic Development; Engineering; Enzymes; Eukaryota; Exploratory/Developmental Grant; Gene Expression; Gene Silencing; Genes; Genome; Goals; Histone Code; Histone H3; Histones; Instruction; Language; Lysine; Malignant Neoplasms; Methods; Methylation; Methyltransferase; Modeling; Modification; Nucleosomes; Phosphorylation; Physiological; Process; Promoter Regions; Proteins; Regulation; Repression; Role; Site; Specificity; Tail; Techniques; Technology; Testing; Ubiquitination; United States National Institutes of Health; Zebrafish; Zinc Fingers; blastomere structure; cell type; demethylation; design; genome-wide; histone methyltransferase; histone modification; in vivo; novel; protein function; tumor

DESCRIPTION (provided by applicant): The long-range goal of the proposed studies is to determine how chromatin modulates gene expression during embryogenesis. The focus is on the role of two histone modifications that are associated with developmental control genes. Trimethylation of lysine 27 of histone H3 (H3K27me3) is commonly associated with silenced genes, whereas trimethylation of lysine 4 of histone H3 (H3K4me3) is commonly associated with genes that are active or permissive to be activated. Studies in pluripotent embryonic cells have revealed that both these marks are associated with developmental control genes before these genes are expressed. These bivalent H3K4me3/H3K27me3 marks have been proposed to contribute to the quantitative, temporal or spatial precision of developmental gene expression, but the field lacks a technology that would allow a direct test of these hypotheses. This project aims to (1) develop a method to induce specific histone modifications at defined genes and (2) apply this technology to determine the role of bivalent histone marks during zebrafish embryogenesis. The method involves the design of zinc finger fusion proteins that have histone methyltransferase or demethylase activities. These proteins will be targeted to defined genes to change H3K4 or H3K27 methylation. The technology will then be used to analyze the consequences of disrupting or introducing H3K4me3 or H3K27me3 marks at specific genes. Since specific histone modifications accompany developmental and physiological states in all eukaryotes, developing a technology for the gene-specific modulation of histone marks would help elucidate the role of such modifications in numerous settings. The proposed technology could also have direct impact in studying and modulating disease processes. For example, abnormal histone modifications have been implicated in several cancers. Local manipulation of histone modifications might help to determine which of the many misregulated genes are responsible for tumor formation. It might even become possible to use the proposed technology to specifically activate disease-protective genes or repress disease-inducing genes. PUBLIC HEALTH RELEVANCE: This project addresses how the myriad of different cell types form during animal development. Abnormal regulation of this process leads to birth defects and cancer, whereas methodical manipulation of this process can help understand and treat degenerative diseases.

描述（由申请人提供）：拟议研究的长期目标是确定染色质如何在胚胎发生过程中调节基因表达。重点是与发育控制基因相关的两种组蛋白修饰的作用。组蛋白 H3 赖氨酸 27 (H3K27me3) 三甲基化通常与沉默基因相关，而组蛋白 H3 赖氨酸 4 (H3K4me3) 三甲基化通常与活跃或允许激活的基因相关。对多能胚胎细胞的研究表明，这两种标记在表达之前都与发育控制基因相关。这些二价 H3K4me3/H3K27me3 标记已被提议有助于发育基因表达的定量、时间或空间精度，但该领域缺乏允许直接测试这些假设的技术。该项目旨在 (1) 开发一种在特定基因上诱导特定组蛋白修饰的方法，以及 (2) 应用该技术确定二价组蛋白标记在斑马鱼胚胎发生过程中的作用。该方法涉及设计具有组蛋白甲基转移酶或去甲基酶活性的锌指融合蛋白。这些蛋白质将靶向特定基因以改变 H3K4 或 H3K27 甲基化。然后，该技术将用于分析在特定基因上破坏或引入 H3K4me3 或 H3K27me3 标记的后果。由于特定的组蛋白修饰伴随着所有真核生物的发育和生理状态，因此开发一种组蛋白标记的基因特异性调节技术将有助于阐明此类修饰在多种环境中的作用。拟议的技术还可能对研究和调节疾病过程产生直接影响。例如，异常的组蛋白修饰与多种癌症有关。组蛋白修饰的局部操作可能有助于确定众多失调基因中哪些基因与肿瘤形成有关。甚至有可能使用所提出的技术来特异性激活疾病保护基因或抑制疾病诱导基因。 公共健康相关性：该项目探讨了动物发育过程中无数不同细胞类型的形成方式。这一过程的异常调节会导致出生缺陷和癌症，而有条理地操纵这一过程可以帮助理解和治疗退行性疾病。