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

• 批准号: 6986005
• 负责人: MARK R PARTHUN
• 金额: $ 21.46万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6986005
• 关键词: chromatin; clinical research; histones; mass spectrometry; tumor suppressor genes

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 必须高度压缩才能包装在细胞核内。这 压缩是通过形成称为染色质的复杂核蛋白结构来实现的。 染色质是一种高度动态的结构，对涉及的细胞过程产生强大的影响 获取染色体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机制 哪个染色质结构受到调节。核心组蛋白已被证明经历多种类型的 修饰如乙酰化、甲基化、磷酸化、泛素化和ADP-核糖基化。在 此外，这些修饰通常出现在核心组蛋白的多个位点。我们的主要关注点 目前的建议是使用灵敏的质谱技术作为基础 组蛋白翻译后修饰的全面鉴定、绘图和表征。 我们在这些研究中的最初目标是使用质谱分析从牛分离的核心组蛋白 胸腺。这将允许识别和绘制完整的组蛋白翻译后谱 高等真核生物染色质中存在的修饰。由于新的修饰位点是 鉴定后，将利用酵母独特的特性，对其体内功能进行表征 遗传操纵该生物体核心组蛋白的能力。 许多观察结果表明，染色质结构的正确调节在 防止癌症不受控制的细胞生长特征。因此，我们建议进行实验 寻求识别与相关的组蛋白修饰程度或模式的特征改变 慢性淋巴细胞白血病。 正如该计划项目的其他部分所述，染色质结构的修饰正在 作为一种潜在的化疗策略进行探索。组蛋白脱乙酰酶的小分子抑制剂 显示出控制癌细胞生长和分化的潜力。为了最有效地利用这些 化合物，更准确地表征诱导的组蛋白修饰的变化至关重要 通过这些药物。因此，我们建议使用质谱来全面表征 用化疗药物治疗白血病细胞导致的组蛋白修饰的变化 目标染色质结构。