Histone acetylation by ethanol:Mechanisms & consequence

乙醇乙酰化组蛋白：机制

• 批准号: 6911646
• 负责人: Shivendra D Shukla
• 金额: $ 20.92万
• 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 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6911646
• 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正在成为转录激活和基因沉默的重要机制。我们最近进行了最初的观察，即乙醇优先引起核组蛋白 H3 第 9 位赖氨酸乙酰化的惊人（8-10 倍）增加，并且这种增加对低至 5 mM 的乙醇非常敏感。我们的长期目标是解决这种乙酰化如何以及为何发生？该项目将奠定基础。我们的假设是“乙醇会在体外和体内引起组蛋白的选择性乙酰化，这将因此影响转录机制。”来自体内乙醇喂养的大鼠的肝细胞和肝脏的原代培养物将作为该项目的主要模型。旨在构建该项目未来发展框架的三个目标。 目的我将确定乙醇诱导肝细胞中组蛋白乙酰化的特征。为此，将研究这种效应的时间过程及其可逆性。我们还将探索不同组织来源的细胞中乙醇诱导的 H3 乙酰化反应的特异性和多样性。 目标 II 将确定肝细胞中组蛋白乙酰化的机制。该方案将检查乙醇代谢抑制剂、单个乙醇代谢物或衍生成分的影响，以及乙酰辅酶A水平变化在乙酰化反应中的作用。该目标还将解决是否涉及组蛋白乙酰转移酶（HAT）表达和/或激活的增加或组蛋白脱乙酰酶（HDAC）的抑制。 目标 III 致力于体内研究，确定液体饮食乙醇喂养对大鼠肝脏 H3 乙酰化的影响。将进行使用染色质免疫沉淀（CHIP）测定研究乙酰化对基因表达的影响的实验，这些实验将构成该项目未来扩展的基础。 这项探索性的 R21 提案涉及酒精领域的新方向，并有望在转录水平上提供乙醇效应的机制数据。此外，参与组蛋白修饰的“机器”将作为开发治疗工具的新目标，用于预防和治疗受乙醇影响的肝脏和其他器官系统中乙醇引起的细胞损伤。