Structure & Expression of Mammal ALC Dehydrogenase Genes

结构

• 批准号: 7887153
• 负责人: HOWARD J EDENBERG
• 金额: $ 3.45万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-03 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7887153
• 关键词: Address; Affect; Alcohol consumption; Alcohol dehydrogenase; Alcoholism; Alcohols; All-Trans-Retinol; Binding; Bioinformatics; Biological Assay; Caucasians; Caucasoid Race; Chromosomes, Human, Pair 4; Code; Consumption; Coupled; Data; Detection; Disease; Distant; Drug or chemical Tissue Distribution; Elements; Enzymes; Ethanol Metabolism; Evolution; Gel; Gene Cluster; Gene Duplication; Gene Expression; Gene Expression Regulation; Gene Family; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Goals; Grant; Human; In Vitro; Individual; Individual Differences; Lead; MHC Class I Genes; Mammals; Metabolic; Metabolism; Nomenclature; Nucleic Acid Regulatory Sequences; Oxidoreductase; Pattern; Pharmacogenetics; Physiological; Population; Protein Binding; Regulation; Research; Research Personnel; Risk; Sequence Analysis; Specificity; Structure; Terminology; Testing; Tissues; Transfection; alcohol risk; member; programs; research study; transcription factor

The long term goals of this research are to understand the mechanisms that regulate humanalcohol dehydrogenase (ADH) gene expression, and the physiological and pathological consequences of alterations in ADH expression. Wehypothesize that differences in the expression of the ADH genes affect the metabolism of alcohol, the risk for alcoholism, and thephysiological and pathological consequences of alcohol consumption. To begin testing this hypothesis we will identify the regions that control ADH expression, identify poly-morphisms in these regions, and then analyze whether these polymorphisms affect gene expression. We will identify cw-acting elements important in the regulation of the human ADH genes, with primary focus on the class I genes and ADH7. These have different patterns of expression, and produce enzymes that influence important metabolic processes including metabolism of ethanol and retinol. Cw-acting elements will be identified by a combination of functional studies (transfections) and protein-binding analyses (DNasel footprintingand gel retardation). We will identify transcription factors that bind to these sequences. The ADH genes are clustered on chromosome 4, and we hypothesize that distant flanking sequences are important in regulating this group of genes. We will test this hypothesis by studying more distant sequences, extending at least 10 kb from the coding regions. We will also analyze the regulation of large chromosomal segments (BACs) carrying groups of ADH genes to detect potential interactions in a chromosomal context. We hypothesize that inter-individual differences in regulatory sequences cause differences in ADH gene expression. We will identify and characterize polymorphisms in the regulatory regions and test their effects on gene expression in vitro. These two aims will be closely coupled, with the localization of cw-acting elements directing our search for polymorphisms, and our detection of functional polymorphisms helping to prioritize further study of the cw-acting elements. These studies will contribute to our understanding of the genetic factors underlyingdifferences among individuals in the metabolic, pharmacological and pathological effects of alcohol consumption. They will also increase our basic understanding of gene regulation.

这项研究的长期目标是了解调节人类酒精的机制 脱氢酶（ADH）基因表达以及改变的生理和病理后果 在ADH表达中。 wehypothes，ADH基因表达的差异会影响 酒精的代谢，酒精中毒的风险以及生理和病理学的后果 饮酒。要开始检验这一假设，我们将确定控制ADH的区域 表达，确定这些区域中的多态性，然后分析这些多态性是否影响 基因表达。 我们将确定在调节人ADH基因中重要的CW作用元件，主要 专注于I类基因和ADH7。这些具有不同的表达方式，并产生酶 影响重要的代谢过程，包括乙醇和视黄醇的代谢。 CW作用元素 将通过功能研究（转染）和蛋白质结合分析的组合来确定 （DNASEL足迹和凝胶阻滞）。我们将确定与这些序列结合的转录因子。 ADH基因聚集在4号染色体上，我们假设遥远的侧翼序列是 在调节这一基因方面重要。我们将通过研究更遥远的序列， 从编码区域延长至少10 kb。我们还将分析大染色体的调节 携带ADH基因组的片段（BAC）在染色体环境中检测潜在相互作用。 我们假设调节序列的个体间差异会导致ADH基因的差异 表达。我们将确定和表征监管区域中的多态性并测试其效果 在体外基因表达上。这两个目标将与CW-Acting的本地化紧密耦合 指导我们寻找多态性的要素，以及我们对功能多态性的检测有助于 优先考虑对CW-COTING元件的进一步研究。 这些研究将有助于我们理解对遗传因素的理解 饮酒的代谢，药理和病理影响的个体。他们会的 还增加了我们对基因调节的基本理解。