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复合物而被压缩。该复合物的主要蛋白质组分是核心组蛋白H2A、H2B、H3和H4。核心组蛋白形成八聚体复合物，DNA围绕该复合物缠绕以形成核小体，核小体是作为高级染色质折叠的基础的基本重复结构。由于DNA包装到染色质中严重限制了DNA的可及性，因此调节染色质结构的能力对于需要接近DNA的细胞过程（例如转录、DNA复制和DNA修复）至关重要。核心组蛋白的翻译后修饰已经成为细胞中染色质结构调节的重要机制。因此，核心组蛋白是许多修饰的位点。在过去几年中出现的组蛋白修饰的一个有趣的方面是，它们并不是孤立地起作用的。现在有几个不同修改之间串扰的例子。例如，组蛋白H3赖氨酸4和79的甲基化需要组蛋白H2B赖氨酸123的泛素化。因此，一个组蛋白上的修饰可以影响其他修饰的存在。我们的建议是结合联合收割机酵母遗传学，稳定同位素标记和质谱，全面和定量地确定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心组蛋白中组蛋白修饰的所有位点。然后，我们将使用稳定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。通过这种方式，将获得核心组蛋白中修饰之间存在的所有串扰的无偏图像。公共卫生相关性：由于其巨大的线性长度，真核生物的基因组DNA需要高度浓缩以适应细胞内部。DNA通过与被称为组蛋白的蛋白质包装而浓缩成一种称为染色质的复合物。细胞还必须能够调节DNA特定区域的浓缩程度，以便在必要时可以访问DNA。调节染色质凝聚的一个重要机制是通过对组蛋白的修饰。该提案旨在确定组蛋白修饰的不同位点之间存在的相互作用。了解组蛋白修饰如何发挥作用对人类健康很重要，因为组蛋白修饰和染色质结构调节的缺陷在癌症等疾病中起着关键作用。