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: 8629759
• 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/8629759
• 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; Genomic DNA; 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; public health relevance; 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.

描述（由申请人提供）：真核细胞每次分裂时，不仅其基因组DNA必须复制，而且其基础染色质结构也必须复制。染色质结构的忠实繁殖对于真核生物基因组的包装和保护以及表观遗传遗传的转录程序的维持是必要的。我们研究计划的主要目标是破译染色质的主要蛋白质组分（核心组蛋白H2A，H2B H3和H3以及连接组蛋白H1）与基因组DNA结合形成染色质的机制。 该提案的具体重点是一类被称为B型组蛋白乙酰转移酶的酶的表征。这些酶负责新合成的组蛋白的翻译后乙酰化。虽然几十年来已经知道新合成的组蛋白在染色质组装过程中被乙酰化，但这些修饰的功能尚不清楚。我们已经提出了一些研究，这将有助于确定B型组蛋白乙酰转移酶Hat 1 p在染色质组装中的作用。 第一个具体目标是利用S。酿酒酵母作为模型系统，研究Hat1p和乙酰化的新合成的组蛋白。我们将使用实验系统，这将使我们能够直接测定Hat1p在几种不同情况下对染色质组装的影响。此外，将使用各种酵母分子遗传技术来破译已在新合成的组蛋白上鉴定的多个乙酰化位点的独特和重叠功能。最后，我们将继续分离和表征一种新的染色质组装因子，我们已经确定在酵母提取物。 第二个具体目标扩展了我们对酵母酶的研究，以表征哺乳动物Hat 1。我们将使用生物化学技术来分离和表征含有人类Hat 1酶的复合物。此外，我们将表征Hat 1小鼠敲除模型，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。