Histone acetylation by ethanol:Mechanisms & consequence

乙醇乙酰化组蛋白：机制

• 批准号: 6754225
• 负责人: Shivendra D Shukla
• 金额: $ 17.28万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6754225
• 关键词: acetyl coA; acetylation; acyltransferase; amidohydrolases; cell nucleus; cell population study; chromatin; chromatin immunoprecipitation; coenzyme A; drug metabolism; enzyme activity; ethanol; gene expression; genetic transcription; histones; kidney cell; laboratory rat; liver cells; neurons; stimulant /agonist; time resolved data; tissue /cell culture; vascular endothelium

DESCRIPTION (provided by applicant): Post-translational modifications in histones are emerging as important mechanisms for the transcriptional activation and gene silencing. We have recently made the original observation that ethanol preferentially caused a spectacular (8-10 fold) increase in acetylation of nuclear histone H3 at Lys 9 and that this increase was remarkably sensitive to as low as 5 mM ethanol. Our long-term goal is to address how and why this acetylation occurs? This project will lay the foundation. Our hypothesis is that "ethanol causes selective acetylation in histones, both in vitro and in vivo, and that this would consequently affect the transcriptional machinery." Primary cultures of hepatocytes and livers from in vivo ethanol fed rats will serve as the main model in this project. There are three aims designed to build the framework for future development of this project. Aim I will determine the characteristics of ethanol induced histone acetylation in hepatocytes. In this aim, time course of this effect and its reversibility will be studied. We will also explore the specificity and diversity of this ethanol induced H3 acetylation response in cells of different tissue origins. Aim II will determine the mechanism(s) of histone acetylation in hepatocytes. The protocols will examine the effects of inhibitors of ethanol metabolism, individual ethanol metabolites or derived components, and the role of changes in levels of acetyl CoA in the acetylation response. This aim will also address whether increased expression and/or activation of histone acetyl transferase (HAT) or an inhibition of histone deacetylase (HDAC) are involved. Aim III is devoted to in vivo studies where effect of liquid diet ethanol feeding on rat liver H3 acetylation will be established. Experiments to study the consequence of acetylation on gene expression using chromatin immunoprecipitation (CHIP) assay will be performed and these experiments will form the basis for future extension of this project. This exploratory R21 proposal deals with a new direction in the alcohol field and offers the promise of providing mechanistic data into the ethanol effects at transcriptional level. Furthermore, "machinery" involved in histone modification will serve as a new target to develop therapeutic tools for prevention and treatment of ethanol induced cellular damage in liver and other organ systems affected by ethanol.

描述（由申请人提供）： 组蛋白的翻译后修饰是作为转录激活和基因沉默的重要机制而出现的。我们最近对乙醇优先引起乙醇在LYS 9处的核组蛋白H3的乙酰化增加，并且这种增加非常敏感到低至5 mM乙醇。我们的长期目标是解决这种乙酰化如何以及为什么发生？该项目将奠定基础。我们的假设是“乙醇在体外和体内引起组蛋白的选择性乙酰化，因此这会影响转录机械。”来自体内乙醇喂养大鼠的肝细胞和肝脏的原发性培养物将作为该项目的主要模型。有三个目标旨在为该项目的未来开发建立框架。 目的我将确定乙醇诱导的肝细胞中乙醇诱导的组蛋白乙酰化的特征。在此目标中，将研究这种效果及其可逆性的时间过程。我们还将探索该乙醇诱导的H3乙酰化反应的特异性和多样性。 AIM II将确定肝细胞中组蛋白乙酰化的机制。这些方案将检查乙醇代谢，单个乙醇代谢物或衍生成分的抑制剂的影响，以及乙酰COA水平变化在乙酰化反应中的作用。该目标还将解决组蛋白乙酰转移酶（HAT）的表达和/或激活增加或组蛋白脱乙酰基酶（HDAC）的抑制。 AIM III致力于体内研究，其中将建立液体饮食乙醇对大鼠肝H3乙酰化的影响。将对使用染色质免疫沉淀（CHIP）测定法研究乙酰化对基因表达的后果的实验，这些实验将构成该项目将来扩展的基础。 该探索性R21提案涉及酒精场中的新方向，并提供了在转录级别提供乙醇效应的机械数据的希望。此外，参与组蛋白修饰的“机械”将成为开发预防和治疗乙醇诱导细胞损伤的肝脏和其他受乙醇影响的细胞损伤的新目标。