Project 3: Mechanisms of Methyltransferase Dysregulation by Oncohistones

项目 3：肿瘤组蛋白甲基转移酶失调的机制

• 批准号: 10024845
• 负责人: Tom Muir
• 金额: $ 27.1万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-09 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10024845
• 关键词: 3-Dimensional; Address; Affect; Alleles; Architecture; Bar Codes; Biochemical; Biochemical Genetics; Biochemical Process; Biological; Biology; Biophysics; Categories; Cell Culture System; Cell Survival; Chemicals; Chemistry; Chromatin; Chromatin Structure; Collaborations; Collection; DNA; Data; Databases; Deposition; Disease; Epigenetic Process; Excision; Funding; Future; Genes; Genetic Screening; Genetic Transcription; Genome; Genomic DNA; Genomic approach; Genomics; Globular Region; Goals; Histone Fold; Histone H3; Histones; Human; Investigation; Lead; Libraries; Malignant Childhood Neoplasm; Malignant Neoplasms; Methods; Methyltransferase; Missense Mutation; Modification; Molecular; Mutation; N-terminal; Nucleosomes; Output; Phase; Phenotype; Physiological; Polymers; Post-Translational Protein Processing; Process; Proteins; Proteomics; Regulation; Role; Screening Result; Screening procedure; Signal Transduction; Structure; Tail; Technology; Testing; Therapeutic; Trans-Activators; Transcriptional Regulation; Validation; Work; Yeasts; base; biophysical properties; cancer cell; cancer type; chromatin remodeling; design; dimer; falls; genome integrity; high throughput screening; in vivo evaluation; knowledge base; member; mouse model; mutant; programs; screening; success; tool; tumor; tumorigenesis; yeast genetics

PROJECT SUMMARY SUMMARY: The broad goal of this project is to study the detailed molecular mechanisms by which histone mutations associated with cancers perturb chromatin states, leading to disease. The next phase of this program is driven by our recent work with the Allis lab revealing a vastly expanded landscape of putative oncohistone mutations - over 4200 missense mutations in dozens of human cancers. Remarkably, the mutations occur in all four of the core histones and in both the N-terminal tails and globular histone fold domains. The very large number of newly identified oncohistones makes experimental characterization daunting. A key challenge is to identify those mutations that act as potential cancer drivers from those that are merely a consequence of the high mutation burden of a given tumor (i.e. passengers). We will work closely with other members of the P01 team to identify those mutations that most likely to fall into the former category and that, as such, merit in vivo testing. Strategically, we will approach this problem using newly developed high-throughput biochemical and yeast genetic screening tools (Aim 1) that are expected to identify mutations that alter chromatin stability and/or that affect the activity of trans-acting factors that operate on the chromatin polymer. Validation of the ‘hits’ from the screening studies will form the core of Aims 2 & 3 where we will combine the use of chemically-defined chromatin templates with biochemical, proteomic and genomic approaches to develop a mechanistic understanding of how select mutations impact chromatin state. In particular, we will study how breakdown in nucleosome symmetry, as a result of sub-stoichiometric incorporation of mutant histones, affects key processes such as chromatin remodeling and transcription. We imagine that many of the technologies developed in the context of this work will have broad utility in the epigenetics field generally. Ultimately, the biochemical knowledge base generated in the course of this program will motivate future therapeutic efforts for treating cancers associated with oncohistone mutants.

项目摘要 本项目的主要目标是研究组蛋白在细胞内表达的详细分子机制。 与癌症相关的突变扰乱染色质状态，导致疾病。这个项目的下一阶段 是由我们最近与Allis实验室的工作推动的，该实验室揭示了推定的癌组蛋白的广泛分布。 突变-在几十种人类癌症中有超过4200种错义突变。值得注意的是，突变发生在所有 四个核心组蛋白和两个N-末端尾部和球状组蛋白折叠域。非常大的 新鉴定的癌组蛋白的数量使得实验表征令人望而生畏。一个关键的挑战是 从那些仅仅是癌症的后果的突变中识别出那些作为潜在癌症驱动因素的突变。 给定肿瘤的高突变负荷（即乘客）。我们将与P01的其他成员密切合作， 研究小组确定了那些最有可能属于前一类的突变，因此，在体内， 试验.从战略上讲，我们将使用新开发的高通量生物化学和 酵母遗传筛选工具（Aim 1），预期可鉴定改变染色质稳定性的突变，和/或 影响对染色质聚合物起作用的反式作用因子的活性。验证来自 筛选研究将构成目标2和目标3的核心，在目标2和目标3中，我们将联合收割机与化学定义的 染色质模板与生物化学，蛋白质组学和基因组学方法，以开发一种机制， 了解选择性突变如何影响染色质状态。特别是，我们将研究如何在 核小体对称性，作为突变组蛋白亚化学计量掺入的结果，影响关键过程 如染色质重塑和转录。我们可以想象，在过去的几十年里， 这项工作的背景通常在表观遗传学领域具有广泛的实用性。最终，生物化学知识 在这个项目过程中产生的基础将激励未来治疗癌症的努力 与癌组蛋白突变体有关。