Histone Chaperones and Acetyltransferases in Chromatin Transitions

描述（由申请人提供）：在每个真核细胞的细胞核内存在一个基本的悖论：遗传物质必须被组织和压缩，同时保持可由转录机制读出。组蛋白是这项任务的核心，最终将染色体DNA压缩数千倍以适应细胞核。基因组可及性可以部分地通过组蛋白伴侣和组蛋白乙酰转移酶（HAT）的作用来实现。组蛋白分子伴侣直接与组蛋白相互作用，并可以在DNA上组装和/或分解它们。HAT共价修饰组蛋白，从而具有改变染色质结构的潜力。这两个项目的研究人员发现了组蛋白伴侣和组蛋白乙酰转移酶活性之间新的令人兴奋的联系。然而，实际上对这些蛋白质共同管理基因组可及性的机制一无所知。这个项目的目标是确定HAT和组蛋白伴侣的结构和功能之间的分子相互关系。这将通过测试三个共同的假设来实现，这允许在所有层面（遗传，生化，代谢，酶和结构）进行多维和深入的调查。在项目1中，我们将研究组蛋白分子伴侣和HAT如何共同发挥作用，从组装成核小体阵列的特定启动子DNA模板中驱逐组蛋白的生化基础。在项目2中，我们将继续进行组蛋白伴侣的理化和结构研究及其对乙酰化染色质的影响。在项目3中，我们将使用体内酵母遗传学和体外生物化学的组合来研究组蛋白伴侣和HAT的遗传和物理相互作用。通过三个研究项目的整合，信息和试剂的交换，以及生物物理学，蛋白质表达和纯化（PEP）和管理核心的重大贡献，该计划产生的结果将深刻影响我们对基因组可及性如何通过调节染色质动力学来调节的理解。 相关性：本提案中所述的研究将提供有关基因组组织和DNA编码信息可及性的重要信息。这些关于基因表达的研究与细胞分裂过程、多细胞生物体的发育和疾病进展（如癌症）直接相关。

DESCRIPTION (provided by applicant): There is a fundamental paradox within the nucleus of every eukaryotlc cell: the genetic material must be organized and compacted, while remaining accessible for readout by the transcription machinery. Histone proteins are at the core of this task and ultimately compact the chromosomal DNA thousands of fold to fit into the nucleus. Genome accessibility can be achieved in part by the actions of histone chaperones and histone acetyltransferases (HATs). Histone chaperones interact directly with histones and can assemble and/or disassemble them on DNA. HATs covalently modify the histones and thereby have the potential to alter chromatin structure. The investigators on this PROGRAM OF PROJECTS have discovered new and exciting linkages between the activities of histone chaperones and histone acetyltransferases. However, virtually nothing is known about the mechanisms by which these proteins work together to manage genome accessibility. The goal of this PROGRAM OF PROJECTS is to determine the molecular interrelationships between the structure and function of HATs and histone chaperones. This will be accomplished by testing three shared hypotheses, which allows for a multi-dimensional and intensive investigation at all levels (genetically, biochemically, thermodynamically, enzymatically, and structurally). In PROJECT 1, we will investigate the biochemical basis for how the histone chaperones and HATs function together to evict histones from a specific promoter DNA template assembled into nucleosomal arrays. In PROJECT 2, we will pursue physicochemical and structural studies related to histone chaperones and their effect on acetylated chromatin. In PROJECT 3, we will use a combination of in vivo yeast genetics and in vitro biochemistry to study the genetic and physical interactions of histone chaperones and HATs. Through the integration of the three research projects, the exchange of information and reagents, and the significant contributions of the Biophysics, Protein Expression and Purification (PEP) and Administrative Cores, the results generated from this Program will profoundly impact our understanding of how genome accessibility is regulated by modulating chromatin dynamics. Relevance: The studies described in this proposal will provide important information about the organization of the genome and accessibility of the information encoded in the DNA. These studies on gene expression are directly relevant to processes of cell division, development of multi-cellular organisms, and disease progression (such as cancer).