Structure/Function of Histone Acetyltransferases

组蛋白乙酰转移酶的结构/功能

• 批准号: 7522931
• 负责人: Ronen Marmorstein
• 金额: $ 35.54万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7522931
• 关键词: ATF2 gene; Acetyl Coenzyme A; Acetylation; Acetyltransferase; Address; Archaea; Binding; Biochemical; Biochemistry; Biological Process; Catalytic Domain; Cell Cycle Progression; Characteristics; Chemistry; Chromatin; Coenzyme A; Collaborations; Colorectal; Complex; Development; Disease; Dosage Compensation (Genetics); EP300 gene; Enzymatic Biochemistry; Enzymes; Family; Funding; Gene Activation; Gene Expression; Genes; Genome Stability; Goals; Histone H3; Histones; Homeostasis; Hormones; Human; Lead; Link; Lysine; Malignant Neoplasms; Mediating; Modeling; Molecular; Molecular Chaperones; Mutagenesis; N-terminal; Nitrogen; Pharmaceutical Preparations; Play; Property; Proteins; Reagent; Recombinants; Regulation; Reporting; Roentgen Rays; Role; Signal Transduction; Structure; Substrate Specificity; Tail; Therapeutic; Universities; Work; X-Ray Crystallography; Yeasts; base; cofactor; design; falls; follow-up; histone acetyltransferase; histone modification; human disease; in vivo; inhibitor/antagonist; insight; leukemia; malignant stomach neoplasm; man; novel; overexpression; public health relevance; small molecule; transcription factor; yeast protein

DESCRIPTION (provided by applicant): The overall goal of this project is to elucidate the structure and function of histone acetyltransferase (HAT) enzymes. These proteins use the acetyl-CoA cofactor to acetylate the 6 nitrogen of specific lysine residues within the N-terminal tails of core histones and work in concert with other histone modification enzymes to provide a mechanistic link between chromatin alteration and gene activation. HAT proteins are found from yeast to man and regulate genes involved in several different biological processes including cell cycle progression, dosage compensation and hormone signaling. Aberrant HAT function has also been correlated with human disease, including leukemic translocations involving the CBP and MOZ HATs, and p300 HAT alterations in colorectal and gastric cancers. HAT proteins fall into subfamilies with characteristic substrate specificity for histones, and in some cases non-histone substrates. Over the last funding periods, we have used X-ray crystallography and complementary biochemical and enzymatic analysis to characterize the substrate binding and catalytic properties of the Gcn5/PCAF, MYST and most recently the p300/CBP subfamilies of HAT proteins. Our results demonstrate that while these different HAT families have a structurally conserved central enzymatic core, they employ different catalytic mechanisms. We also show that the regions within the HAT domain that flank the catalytic core within these enzymes have divergent structure and function and are thus particularly important in imparting different transcriptional regulatory properties to these HAT enzymes. We will now continue to use a combination of structure, biochemistry and enzymology to carry out additional studies on the MYST and p300/CBP family of HATs and to extend our studies to 2 other acetyltransferases, an evolutionarily ancient archaeal HAT called PAT and a newly characterized yeast HAT called Rtt109 that cooperates with the histone chaperone protein Vps75 to acetylate K56 of histone H3 to promote genomic stability. Specifically, the four aims of the proposal are to (1) Study the mechanism of p300/CBP regulation by an autoregulatory loop and by the ATF2 transcription factor, and screen for p300/CBP inhibitors; (2) Characterize the mechanism for acetylation, substrate recognition and autoregulation by the archaeal PAT acetyltransferase; (3) Characterize the mechanism for acetylation by the Rtt109 HAT and its modulation by the Vps75 histone chaperone; and (4) Investigate the catalytic mechanism of the MYST HATs and characterize the structure and function of the heterotrimeric Sas2/4/5 complex. Together, these studies will provide new molecular insights into the substrate specificity and catalytic mechanism of histone acetyltransferases, and how these activities are modulated by intra-auto-regulatory domains and other associated protein factors. PUBLIC HEALTH RELEVANCE: Statement Histone Acetyltranferase (HAT) enzymes play key roles in regulating gene expression and aberrant HAT function has also been correlated with several human diseases, including leukemic translocations involving the CBP and MOZ HATs, and p300 HAT alterations in colorectal and gastric cancers. The overall goal of this project is to elucidate the structure, chemistry and function of histone acetyltransferase (HAT) enzymes and to use this information to aid in the design small molecule drugs to treat HAT-associated diseases.

描述（由申请人提供）：该项目的总体目标是阐明组蛋白乙酰转移酶（HAT）的结构和功能。这些蛋白质使用乙酰辅酶 A 辅因子乙酰化核心组蛋白 N 末端尾部特定赖氨酸残基的 6 个氮，并与其他组蛋白修饰酶协同工作，在染色质改变和基因激活之间提供机制联系。 HAT 蛋白从酵母到人类都有发现，可调节参与多种不同生物过程的基因，包括细胞周期进程、剂量补偿和激素信号传导。 HAT 功能异常也与人类疾病相关，包括涉及 CBP 和 MOZ HAT 的白血病易位，以及结直肠癌和胃癌中的 p300 HAT 改变。 HAT 蛋白属于具有组蛋白底物特异性的亚家族，在某些情况下还具有非组蛋白底物。在过去的资助期间，我们使用 X 射线晶体学和补充生化和酶分析来表征 Gcn5/PCAF、MYST 以及最近的 HAT 蛋白 p300/CBP 亚家族的底物结合和催化特性。我们的结果表明，虽然这些不同的 HAT 家族具有结构保守的中心酶核心，但它们采用不同的催化机制。我们还表明，这些酶内催化核心侧翼的 HAT 结构域内的区域具有不同的结构和功能，因此对于赋予这些 HAT 酶不同的转录调控特性特别重要。现在，我们将继续结合结构、生物化学和酶学对 MYST 和 p300/CBP 家族 HAT 进行更多研究，并将我们的研究扩展到其他 2 种乙酰转移酶，即一种进化上古老的古菌 HAT（称为 PAT）和一种新鉴定的乙酰转移酶。酵母 HAT 称为 Rtt109，它与组蛋白伴侣蛋白 Vps75 配合乙酰化组蛋白 H3 的 K56，以促进基因组稳定性。具体来说，该提案的四个目标是（1）研究自动调节环和ATF2转录因子对p300/CBP的调节机制，并筛选p300/CBP抑制剂； (2) 表征古菌 PAT 乙酰转移酶的乙酰化、底物识别和自动调节机制； (3) 表征 Rtt109 HAT 的乙酰化机制及其受 Vps75 组蛋白伴侣的调节； (4)研究MYST HAT的催化机制并表征异源三聚体Sas2/4/5复合物的结构和功能。总之，这些研究将为组蛋白乙酰转移酶的底物特异性和催化机制，以及这些活性如何通过内部自动调节域和其他相关蛋白质因子调节提供新的分子见解。公共健康相关性：声明组蛋白乙酰转移酶 (HAT) 在调节基因表达中发挥关键作用，异常的 HAT 功能也与多种人类疾病相关，包括涉及 CBP 和 MOZ HAT 的白血病易位，以及结直肠癌和胃癌中的 p300 HAT 改变。该项目的总体目标是阐明组蛋白乙酰转移酶 (HAT) 的结构、化学和功能，并利用这些信息帮助设计治疗 HAT 相关疾病的小分子药物。