Role of Set2 and H3 methylation in chromatin function

Set2 和 H3 甲基化在染色质功能中的作用

基本信息

批准号：
7613459
负责人：
Brian D Strahl
金额：
$ 9.89万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-05-01 至 2009-12-27
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7613459
关键词：
Acetylation Affect Affinity Chromatography Antibodies Binding Biochemical Biochemistry Biological Assay C-terminal Cell Survival Chromatin Chromatin Fiber Chromatin Structure Chromatography Co-Immunoprecipitations Complex DNA DNA Microarray Chip Data Disease Enzymes Event Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Genetic Screening Genetic Transcription Glucose Goals Health Histone H3 Histones Human Immunology In Vitro LIM Domain Lysine Maps Mass Spectrum Analysis Mediating Methylation Methyltransferase Methyltransferase Gene Modeling Modification Mutation Outcome Pathway interactions Peptides Phosphorylation Play Post-Translational Protein Processing Process Protein Binding Protein Binding Domain Proteins Recruitment Activity Regulation Repression Research Personnel Role SET Domain Site Site-Directed Mutagenesis Stream Yeasts base chromatin immunoprecipitation gene repression histone methyltransferase histone modification in vivo insight mutant novel prevent programs promoter protein protein interaction research study

项目摘要

DESCRIPTION (provided by applicant): Regulation of eukaryotic gene expression involves the precise control of chromatin structure. While the manipulation of the chromatin fiber to create and maintain distinct functional domains is poorly understood, histone post-translational modifications are clearly central to this process. The long-range objective of this proposal is to elucidate roles of histone methylation in chromatin function. To achieve this, we will characterize the functions of two independent histone methyltransferase (HMT) activities recently identified in yeast. One of these activities is Set2, which is the sole enzyme in yeast responsible for methylating histone H3 on lysine 36 (K36), a site of modification conserved from yeast to humans. New evidence suggests that Set2-mediated H3 K36 methylation participates in glucose repression, but little else is known about this enzyme or its methylation site. Given recent insights into the roles of histone methylation, we hypothesize that these enzymes regulate gene expression, in part, through the recruitment of down-stream regulatory factors to their cognate methylation sites in chromatin. To that end, we will use a combination of biochemistry, genetics and immunology to define the domains in Set2 responsible for gene repression and histone methylation, characterize Set2's interacting partners, identify proteins that bind K36-methylated H3 and determine the extent to which H3 K36 methylation functions with other histone modifications to regulate the transcription of a gene affected by Set2. We will also determine other chromosomal regions and cellular pathways affected by Set2/H3 K36 methylation in an effort to understand the global functions of this enzyme. The second HMT activity is not yet identified but has been highly purified by chromatography. We will identify this enzyme by biochemical approaches and further characterize its site(s) of methylation and determine the extent to which they occur in chromatin. Since histone modifications play a vital role in chromatin-based functions, advances in our understanding of the enzymes that mediate them, as well as their functional significance, is likely to have a major impact on issues related to human health and disease.

描述（由申请人提供）：真核基因表达的调节涉及染色质结构的精确控制。虽然对染色质纤维创建和维持不同功能域的操纵知之甚少，但组蛋白的翻译后修饰显然是该过程的核心。该建议的远程目标是阐明组蛋白甲基化在染色质功能中的作用。为此，我们将表征最近在酵母中发现的两个独立组蛋白甲基转移酶（HMT）活性的功能。这些活性之一是SET2，它是酵母中负责甲基化组蛋白H3（K36）上的唯一酶，这是一个从酵母到人类的修饰部位。新的证据表明，SET2介导的H3 K36甲基化参与了葡萄糖抑制，但对此酶或其甲基化位点知之甚少。鉴于最近对组蛋白甲基化作用的见解，我们假设这些酶通过募集下游调节因子来调节基因表达，从而调节染色质中的认知甲基化位点。为此，我们将使用生物化学，遗传学和免疫学的组合来定义负责基因抑制和组蛋白甲基化的SET2中的域，表征SET2的相互作用伴侣，鉴定蛋白质的蛋白质结合K36-甲基化的H3并确定H3 K36甲基化与其他组合构型构成基因的构成依赖于ANEN的conee conee conee nege的依赖的程度。我们还将确定受SET2/H3 K36甲基化影响的其他染色体区域和细胞途径，以了解该酶的全球功能。第二个HMT活性尚未确定，但已通过色谱高度纯化。我们将通过生化方法鉴定这种酶，并进一步表征其甲基化的位点，并确定它们在染色质中发生的程度。由于组蛋白修饰在基于染色质的功能中起着至关重要的作用，因此我们对介导其介导的酶以及其功能意义的理解的进步可能对与人类健康和疾病有关的问题产生重大影响。