Identification of Histone Modification Interaction Networks

组蛋白修饰相互作用网络的鉴定

• 批准号: 7570472
• 负责人: Michael A. Freitas
• 金额: $ 26.25万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-20 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7570472
• 关键词: ADP ribosylation; Acetylation; Affect; Antibodies; Biotinylation; Cell physiology; Cells; Chromatin; Chromatin Structure; Collection; Complex; DNA; DNA Packaging; DNA Repair; DNA biosynthesis; Defect; Disease; Eukaryota; Foundations; Genetic Transcription; Genomics; Grant; Health; Higher Order Chromatin Structure; Histone Fold; Histone H2A; Histone H2B; Histone H3; Histones; Human; Isotope Labeling; Length; Lysine; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Methylation; Modification; Mutate; Mutation; N-terminal; Nucleoproteins; Nucleosomes; Phosphorylation; Physical condensation; Play; Positioning Attribute; Post-Translational Modification Site; Post-Translational Protein Processing; Process; Proteins; Reagent; Regulation; Relative (related person); Reliance; Role; Site; Stable Isotope Labeling; Structure; Tail; Yeasts; histone modification; mutant; physical property; public health relevance; yeast genetics

DESCRIPTION (provided by applicant): Eukaryotic DNA is compacted through the formation of a nucleoprotein complex known as chromatin. The primary protein components of this complex are the core histones H2A, H2B, H3 and H4. The core histones form an octameric complex around which DNA wraps to form the nucleosome which is the basic repeating structure that serves as the foundation for higher order chromatin folding. As the packaging of DNA into chromatin places severe constraints on the accessibility of DNA, the ability to regulate chromatin structure is critical for cellular processes that require access to DNA such as transcription, DNA replication and DNA repair. The post-translational modification of the core histones has emerged as an important mechanism by which chromatin structure is modulated in cells. Accordingly, the core histones are the sites of numerous modifications. An intriguing aspect of histones modifications, which has emerged in the past few years, is that they do not function in isolation. There are now several examples of cross-talk between different modifications. For example, the ubiquitylation of histone H2B lysine 123 is required for the methylation of histone H3 lysines 4 and 79. Hence, modifications on one histone can influence the presence of other modifications. Our proposal is to combine yeast genetics, stable isotope labeling and mass spectrometry to comprehensively and quantitatively identify the network of interactions that exist between histone modifications. We will systematically mutate all sites of histone modification in the yeast core histones. We will then use stable isotope labeling and mass spectrometry to quantitate all of the histone modifications in the mutant strain relative to a wild type control. In this way, an unbiased picture will be obtained of all the cross-talk that exists between modifications in the core histones. PUBLIC HEALTH RELEVANCE: Due to its enormous linear length, the genomic DNA of eukaryotes needs to be highly condensed to fit inside cells. The DNA is condensed through packaging with proteins known as histones into a complex called chromatin. Cells must also be able to regulate the degree to which specific regions of DNA are condensed so that the DNA can become accessible when necessary. One important mechanism to regulate chromatin condensation is through modifications to the histone proteins. This proposal seeks to identify interactions that exist between different sites of histone modification. An understanding of how histone modifications function is important for human health as defects in the regulation of histone modifications and chromatin structure play critical roles in diseases such as cancer.

描述（由申请人提供）： 真核DNA通过形成称为染色质的核蛋白络合物来压缩。该复合物的主要蛋白质成分是核心组蛋白H2a，H2b，H3和H4。核心组蛋白形成一个八聚体复合物，DNA围绕它形成核小体，这是基本的重复结构，是高阶染色质折叠的基础。当将DNA包装到染色质中时，将严重的限制在DNA的可及性上，调节染色质结构的能力对于需要访问DNA的细胞过程至关重要，这些过程需要转录，DNA复制和DNA修复。核心组蛋白的翻译后修饰已成为一种重要机制，通过在细胞中调节染色质结构。因此，核心组蛋白是许多修改的位置。过去几年中出现的组蛋白修饰的一个有趣的方面是它们不隔离起来。现在有几个不同修改之间的串扰示例。例如，组蛋白H2b赖氨酸123的泛素化是组蛋白H3赖氨酸4和79所必需的。因此，一种组蛋白的修饰会影响其他修饰的存在。我们的建议是结合酵母遗传学，稳定的同位素标记和质谱法，以全面，定量地识别组蛋白修饰之间存在的相互作用网络。我们将系统地突变酵母核组蛋白中组蛋白修饰的所有位点。然后，我们将使用稳定的同位素标记和质谱法来定量相对于野生型对照的突变株中的所有组蛋白修饰。这样，将获得无偏见的图片，从核心组蛋白中的修饰之间存在的所有串扰。公共卫生相关性：由于其巨大的线性长度，需要高度凝结真核生物的基因组DNA才能适应细胞内部。通过与称为组蛋白的蛋白质包装将DNA凝结成一种称为染色质的复合物。细胞还必须能够调节对DNA的特定区域凝结的程度，以便必要时DNA可以访问。调节染色质冷凝的一种重要机制是通过对组蛋白的修饰。该建议旨在确定组蛋白修饰不同位点之间存在的相互作用。对组蛋白修饰的功能的理解对于人类健康非常重要，因为调节组蛋白修饰和染色质结构在癌症等疾病中起关键作用。