Type B Histone Acetyltransferases and the Assembly of Chromatin Structure

B 型组蛋白乙酰转移酶和染色质结构的组装

• 批准号: 8115669
• 负责人: MARK R PARTHUN
• 金额: $ 31.42万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8115669
• 关键词: Acetylation; Animal Model; Area; Biochemical; Biological; Biological Assay; Biological Models; Biological Process; Birth; Categories; Cell Nucleus; Cells; Chromatin; Chromatin Modeling; Chromatin Structure; Complex; Coupled; DNA; DNA Double Strand Break; Data; Defect; Deposition; Disease; Enzymes; Eukaryotic Cell; Gene Deletion; Generations; Genetic Models; Genetic Techniques; Genome; Genomics; Goals; Histone Acetylation; Histone H1; Histone H1(s); Histone H2A; Histone H3; Histone H4; Histones; Human; Inherited; Investigation; Knockout Mice; Laboratories; Length; Link; Lysine; Maintenance; Mammalian Cell; Modeling; Modification; Molecular Chaperones; Molecular Genetics; Mus; Nuclear; Organism; Pathway interactions; Play; Process; Property; Proteins; RNA; Regulation; Research; Role; Saccharomyces cerevisiae; Series; Site; Specificity; Structure; System; Techniques; Testing; Time; Transcriptional Regulation; Work; Yeasts; base; histone acetyltransferase; human disease; interest; member; novel; programs; protein complex; recombinational repair; research study

DESCRIPTION (provided by applicant): Each time a eukaryotic cell divides it must duplicate, not only its genomic DNA, but the underlying chromatin structure, as well. The faithful propagation of chromatin structure is necessary for the packaging and protection of the eukaryotic genome as well as for the maintenance of epigenetically inherited transcriptional programs. The primary goal of our research program is to decipher the mechanisms by which the primary protein components of chromatin (the core histones H2A, H2B H3 and H3 and the linker histone H1) are brought together with genomic DNA to form chromatin. The specific focus of this proposal is the characterization of a class of enzymes known as type B histone acetyltransferases. These enzymes are responsible for the post-translational acetylation of newly synthesized histones. While it has been known for decades that newly synthesized histones are acetylated during the process of chromatin assembly, the function of these modifications is not known. We have proposed a number of studies that will help to identify the role of the type B histone acetyltransferase Hat1p in chromatin assembly. The first specific aim utilizes S. cerevisiae as a model system to study Hat1p and the acetylation of newly synthesized histones. We will use experimental systems that will allow us to directly assay for the effect of Hat1p on chromatin assembly in several different contexts. In addition, will use a variety of yeast molecular genetic techniques to decipher the unique and overlapping functions of the multiple sites of acetylation that have been identified on newly synthesized histones. Finally, we will continue the isolation and characterization of a novel chromatin assembly factor that we have identified in yeast extracts. The second specific aim extends our studies of the yeast enzyme, to the characterization of mammalian Hat1. We will use biochemical techniques to isolate and characterize complexes containing the human Hat1 enzyme. In addition, we will characterize a Hat1 mouse knockout model in order to identify the function of this type B histone acetyltransferase in a complex organism. PUBLIC HEALTH RELEVANCE: Eukaryotic cells contain an enormous linear length of DNA that must be highly condensed to be packaged inside cells. This packaging results from the formation of a structure known as chromatin that plays an important role in regulating most processes that occur in the nucleus. The importance of chromatin structure is evidenced by the numerous examples of defects in chromatin structure that cause serious human diseases. This proposal seeks to understand the mechanisms by which chromatin is assembled and regulated which will aid in our understanding and treatment of these diseases.

描述(申请人提供)：真核细胞每次分裂时，不仅要复制它的基因组DNA，还要复制潜在的染色质结构。染色质结构的忠实繁殖对于真核基因组的包装和保护以及维持表观遗传的转录程序是必要的。我们研究计划的主要目标是破译染色质的主要蛋白质成分(核心组蛋白H_2A、H_2B、H_3和H_3以及连接物组蛋白H_1)与基因组DNA结合形成染色质的机制。这项建议的具体焦点是一类被称为B型组蛋白乙酰转移酶的酶的特征。这些酶负责新合成的组蛋白的翻译后乙酰化。虽然新合成的组蛋白在染色质组装过程中被乙酰化已经知道几十年了，但这些修饰的功能尚不清楚。我们已经提出了一些研究，这些研究将有助于确定B型组蛋白乙酰转移酶Hat1p在染色质组装中的作用。第一个特定目的是利用酿酒酵母作为模型系统来研究Hat1p和新合成的组蛋白的乙酰化。我们将使用实验系统，使我们能够在几个不同的环境中直接分析Hat1p对染色质组装的影响。此外，还将使用各种酵母分子遗传学技术来破译新合成的组蛋白上已确定的多个乙酰化位点的独特和重叠功能。最后，我们将继续分离和鉴定酵母提取物中发现的一种新的染色质组装因子。第二个特定目的是将我们对酵母酶的研究扩展到哺乳动物Hat1的特征。我们将使用生化技术来分离和鉴定含有人类Hat1酶的复合体。此外，我们将对Hat1小鼠基因敲除模型进行表征，以确定这种B型组蛋白乙酰转移酶在复杂生物体中的功能。 与公共卫生相关：真核细胞含有巨大的线性长度的DNA，必须高度浓缩才能包装在细胞内。这种包装是由一种称为染色质的结构形成的，该结构在调节发生在细胞核中的大多数过程中发挥着重要作用。大量染色质结构缺陷导致人类严重疾病的例子证明了染色质结构的重要性。这项建议旨在了解染色质的组装和调节机制，这将有助于我们对这些疾病的理解和治疗。