Impact of ethanol-induced protein nitration on the histone modification code

乙醇诱导的蛋白质硝化对组蛋白修饰代码的影响

基本信息

批准号：
8638358
负责人：
Stanley M Stevens
金额：
$ 17.45万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-02-15 至 2016-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8638358
关键词：
Acetylation Active Sites Acute Affect Alcohol consumption Alcoholic Liver Diseases Alcohols Animal Model Bioinformatics Biological Assay Cardiovascular Diseases Cell Culture Techniques Cells Charge Chronic Code Custom Data Dependency Development Disease Disease Outcome Dose Drug Targeting Environment Enzymes Epigenetic Process Ethanol Etiology Exposure to Gene Expression Process Generations Genetic Transcription Goals Hepatic Hepatitis Hepatocyte Histone Deacetylase Histone H2A Histone H3 Histones Human In Vitro Incubators Injury Laboratories Lead Light Link Liver Malignant Neoplasms Mass Spectrum Analysis Mediating Modeling Modification Molecular Molecular Weight Mus Neurodegenerative Disorders Nitrates Nitric Oxide Nuclear Nuclear Protein Nuclear Proteins Oxidants Oxidative Stress Peptides Peroxonitrite Pharmacotherapy Phosphorylation Phosphotyrosine Post-Translational Protein Processing Prevention Process Protein Isoforms Proteins Proteomics Publishing Rattus Relative (related person)Research Role Severities Site Specificity Stable Isotope Labeling Stress System Testing Time Tissues Transcription Process Tyrosine Tyrosine Phosphorylation Uric Acid Validation Western Blotting alcohol exposure base cell type epigenetic marker histone modification human NOS2A protein in vivo inhibitor/antagonist link protein liver injury mortality multicatalytic endopeptidase complex multiple reaction monitoring nitration novel public health relevance tool

项目摘要

DESCRIPTION (provided by applicant): Increase in protein tyrosine nitration (PTN) has been associated with numerous diseases and mounting evidence suggests this modification could be a causative factor rather than just a general outcome of disease. The generation of PTN has been implicated in early alcohol-induced hepatitis in mice where inducible nitric oxide synthase (iNOS) and the nitric oxide-derived product, peroxynitrite (a potent agent that causes PTN in the cell), have been identified as major players in the development of this pathophysiological condition of the liver. In an attempt to identify potential hepatic PTN targets, preliminary proteomic studies were performed in our laboratory and the results suggest that histone proteins undergo selective nitration in primary human hepatocyte culture after acute ethanol exposure. Accordingly, the goal of this R21 project is to examine the impact of ethanol-induced PTN on the histone modification code by testing the hypotheses that 1) protein nitration serves as a novel histone modification that could result in subsequent alteration of gene transcription processes and 2) oxidative/nitrative modifications of enzymes that directly modulate histone modification levels could have an impact on various ethanol-induced epigenetic markers (for example, acetylation of histone H3 at Lys9). In Specific Aim 1 of this proposal, we will characterize the process of ethanol-induced tyrosine nitration in histone H2A, H2B, H3, and H4 isoforms. In Specific Aim 2, we will characterize the nuclear nitroproteome after ethanol exposure to identify nitrated proteins associated with altering the histone modification code (for example, HAT or HDAC enzymes). Additionally, our findings from in vitro studies will be validated in an in vivo animal model of chronic ethanol exposure. The results from this project will help shed light on the role of oxidative/nitrative stress in ethanol-induced alterations of the histone modification code and provide further evidence to the link of PTN and disease causation. Identification of nuclear nitration targets and enzyme processes that affect PTN levels will likely provide new information on potential targets for drug therapy in ethanol-induced tissue injury where PTN is linked to injury onset or severity.

描述（由申请人提供）：蛋白质酪氨酸硝化（PTN）的增加与许多疾病相关，并且越来越多的证据表明这种修饰可能是一个致病因素，而不仅仅是疾病的一般结果。 PTN 的产生与早期酒精诱发的小鼠肝炎有关，其中诱导型一氧化氮合酶 (iNOS) 和一氧化氮衍生产物过氧亚硝酸盐（一种在细胞中引起 PTN 的有效物质）已被确定为主要参与者肝脏病理生理状况的发展。为了确定潜在的肝脏 PTN 靶标，我们实验室进行了初步蛋白质组学研究，结果表明，在急性乙醇暴露后，原代人肝细胞培养物中的组蛋白会发生选择性硝化。因此，该 R21 项目的目标是通过测试以下假设来检查乙醇诱导的 PTN 对组蛋白修饰代码的影响：1) 蛋白质硝化作为一种新的组蛋白修饰，可能导致基因转录过程的后续改变，2) ) 直接调节组蛋白修饰水平的酶的氧化/硝基修饰可能对各种乙醇诱导的表观遗传标记（例如组蛋白 H3 在 Lys9 处的乙酰化）产生影响。在本提案的具体目标 1 中，我们将描述组蛋白 H2A、H2B、H3 和 H4 亚型中乙醇诱导的酪氨酸硝化过程。在具体目标 2 中，我们将表征乙醇暴露后的核硝基蛋白质组，以鉴定与改变组蛋白修饰代码（例如 HAT 或 HDAC 酶）相关的硝化蛋白质。此外，我们的体外研究结果将在慢性乙醇暴露的体内动物模型中得到验证。该项目的结果将有助于阐明氧化/硝化应激在乙醇诱导的组蛋白修饰代码改变中的作用，并为 PTN 与疾病因果关系的联系提供进一步的证据。确定影响 PTN 水平的核硝化靶点和酶过程可能会为乙醇引起的组织损伤的药物治疗的潜在靶点提供新信息，其中 PTN 与损伤发生或严重程度相关。