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

基本信息

批准号：
8437176
负责人：
MARK R PARTHUN
金额：
$ 30.32万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
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/8437176
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 public health relevance recombinational repair research study

项目摘要

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都必须复制，不仅复制其基因组 DNA，还复制其底层的染色质结构。染色质结构的忠实传播对于真核基因组的包装和保护以及表观遗传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主要蛋白质成分（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。虽然几十年来人们就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。我们提出了多项研究，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。第一个具体目标利用酿酒酵母作为模型系统来研究 Hat1p 和新合成组蛋白的乙酰化。我们将使用实验系统，使我们能够在几种不同的情况下直接测定 Hat1p 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新合成的组蛋白上已鉴定的多个乙酰化位点的独特且重叠的功能。最后，我们将继续分离和表征我们在酵母提取物中鉴定出的新型染色质组装因子。第二个具体目标将我们对酵母酶的研究扩展到哺乳动物 Hat1 的表征。我们将使用生化技术来分离和表征含有人类 Hat1 酶的复合物。此外，我们将表征 Hat1 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。