Histone Modification and Changes in Chromatin: Silencing of Tumor Suppressor Gene

组蛋白修饰和染色质变化：肿瘤抑制基因的沉默

基本信息

批准号：
7478440
负责人：
MARK R PARTHUN
金额：
$ 32.72万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7478440
关键词：
ADP ribosylation Acetylation Antibodies Bos taurus Cattle Cell Nucleus Cell physiology Cells Characteristics Chromatin Chromatin Structure Chronic Lymphocytic Leukemia Complex DNA Development Differentiation and Growth Eukaryota Eukaryotic Cell Foundations Genetic Genomics Histone Deacetylase Histone H2A Histones Malignant Neoplasms Maps Mass Spectrum Analysis Methylation Modeling Modification Modification Type Nature Nucleoproteins Numbers Organism Pattern Pharmaceutical Preparations Phosphorylation Play Post-Translational Protein Processing Proteins Reagent Recording of previous events Regulation Role Saccharomyces cerevisiae Site Structure System Techniques Thymus Gland Tissue Sample Tumor Suppressor Genes Tumor Tissue Ubiquitination Yeasts base cancer cell differentiation cell growth regulation chemotherapeutic agent in vivo inhibitor/antagonist leukemia novel prevent programs research study small molecule tissue culture tumorigenesis uncontrolled cell growth

项目摘要

The genomic DNA of eukaryotes must be highly compacted for packaging within the nuclei of cells. This compaction is achieved through the formation of a complex nucleoprotein structure known as chromatin. Chromatin is a highly dynamic structure that exerts a powerful influence over cellular processes that involve accessing chromosomal DNA. The primary protein components of chromatin are the core histones H2A, H2B, H3 and H4. The post-translational modification of the core histones is an important mechanism by which chromatin structure is regulated. The core histones have been shown to undergo several types of modification such as acetylation, methylation, phosphorylation, ubiquitination and ADP-ribosylation. In addition, these modifications are often found at multiple sites in the core histones. The main focus of our current proposal is to use the sensitive technique of mass spectrometry as a foundation for the comprehensive identification, mapping and characterization of histone post-translational modifications. Our initial aim in these studies is to use mass spectrometry to analyze core histones isolated from bovine thymus. This will allow for the identification and mapping of the complete spectrum of histone posttranslational modifications present in higher eukaryotic chromatin. As novel sites of modification are identified, the characterization of their in vivo function will be done in yeast, taking advantage of the unique ability to genetically manipulate the core histones in this organism. Many observations point to the critical role that the proper regulation of chromatin structure plays in preventing the uncontrolled cell growth characteristic of cancer. Therefore, we propose experiments that seek to identify characteristic alterations in the extent or pattern of histone modifications that correlate with Chronic Lymphocytic Leukemia. As described in other sections of this program project, the modification of chromatin structure is being explored as a potential chemotherapeutic strategy. Small molecule inhibitors of histone deacetylases have shown potential in controlling the growth and differentiation of cancer cells. To most effectively utilize these compounds, it is vital to more precisely characterize the alterations in histone modifications that are induced by these drugs. Therefore, we propose to use mass spectrometry to comprehensively characterize the changes in histone modification that result from the treatment of leukemia cells with chemotherapeutic agents that target chromatin structure.

真核生物的基因组DNA必须高度紧凑，以在细胞核中包装。这通过形成称为染色质的复杂核蛋白结构来实现压实。染色质是一种高度动态的结构，对涉及细胞过程产生强大的影响进入染色体DNA。染色质的主要蛋白质成分是核心组蛋白H2a， H2B，H3和H4。核心组蛋白的翻译后修饰是一个重要的机制哪种染色质结构受到调节。核心组蛋白已显示出几种类型的修饰，例如乙酰化，甲基化，磷酸化，泛素化和ADP-核糖基化。在此外，这些修饰通常在核心组蛋白的多个位点发现。我们的主要重点当前的建议是使用质谱敏感技术作为基础组蛋白后翻译后修饰的全面识别，映射和表征。在这些研究中，我们最初的目的是使用质谱法分析从牛分离的核心组蛋白胸腺。这将允许识别和映射整体后组蛋白的完整光谱更高的真核染色质中存在的修饰。作为新颖的修改场所是鉴定出来，其体内功能的表征将在酵母中进行，利用独特的优势能够操纵这种生物体的核心组蛋白的能力。许多观察表明，正确调节染色质结构的关键作用是防止癌症不受控制的细胞生长特征。因此，我们提出了实验寻求在与与之相关的组蛋白修饰的范围或模式下识别特征变化慢性淋巴细胞性白血病。如该程序项目的其他部分所述，染色质结构的修饰正在作为一种潜在的化学治疗策略探讨。组蛋白脱乙酰基酶的小分子抑制剂具有显示了控制癌细胞生长和分化的潜力。最有效地利用这些化合物，更精确地表征被诱导的组蛋白修饰的改变至关重要通过这些药物。因此，我们建议使用质谱法全面表征通过化学治疗剂治疗白血病细胞导致组蛋白修饰的变化该靶染色质结构。