SELENIUM REGULATION OF THE GLUTATHIONE PEROXIDASE GENE

硒对谷胱甘肽过氧化物酶基因的调节

基本信息

批准号：
2141850
负责人：
MERRILL CHRISTENSEN
金额：
$ 10.19万
依托单位：
BRIGHAM YOUNG UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-04-01 至 1995-12-31
项目状态：
已结题

项目摘要

Exploration of the mechanisms by which nutrient intake modifies gene expression may reveal the mean by which dietary factors can alter susceptibility to cancer. This work will examine the expression and regulation of the gene for the liver selenoenzyme glutathione peroxidase (GSH:H2O2 oxidoreductase, EC 1.11.19) (GSH-Px). Dependence of that regulation on dietary intake of selenium (Se) will be determined. This requires the isolation and structural and organizational characterization of the rat liver GSH-Px gene. Dependence of transcriptional or translational regulation on diet will be examined by comparing run-off transcripts, accumulation and stability of GSH-Px mRNA, and accumulation and stability of anti-GSH-Px antibody-reactive protein from livers of rats fed Se-deficient or Se-adequate diets. These experiments will show which reactions in the expression of the GSH-Px gene are most sensitive to dietary Se intake. To examine one mechanism by which dietary SE intake may regulate expression of the GSH-Px gene, electrophoretic mobility shift (EMS) assays and methylation interference assays will be done. These experiments will demonstrate binding of trans acting factors (proteins) to sequences in the flanking regions of the GSH-Px gene. Assays performed with lever nuclear extracts from se-adequate and Se-deficient animals will demonstrate the dependence of potential regulatory proteins on dietary Se intake. The use of nuclear extracts in these assays from livers of rats injected with 75 Se will show whether or not selenoproteins themselves, or Se-binding proteins, inducible by dietary Se intake, play a role in the regulation of the GSH-Px gene.

营养素摄入改变基因机制的探索表达可能揭示饮食因素改变的平均值癌症易感性。这项工作将检查表达和肝脏硒酶谷胱甘肽过氧化物酶基因的调控（GSH：H2O2 氧化还原酶，EC 1.11.19）（GSH-Px）。依赖那个将确定硒（Se）膳食摄入量的规定。这需要隔离以及结构和组织大鼠肝脏 GSH-Px 基因的表征。依赖性饮食的转录或翻译调节将通过以下方式进行检查比较 GSH-Px mRNA 的径流转录本、积累和稳定性，抗GSH-Px抗体反应蛋白的积累和稳定性来自喂食缺硒或硒充足饮食的大鼠的肝脏。这些实验将显示 GSH-Px 基因表达中的哪些反应对饮食中硒的摄入量最敏感。研究膳食 SE 摄入量调节表达的一种机制 GSH-Px 基因、电泳迁移率变动 (EMS) 分析和将进行甲基化干扰测定。这些实验将证明反式作用因子（蛋白质）与序列中的结合 GSH-Px 基因的侧翼区域。用杠杆核进行测定来自硒充足和缺硒动物的提取物将证明潜在调节蛋白对膳食硒摄入量的依赖性。用途这些测定中来自注射 75 Se 的大鼠肝脏的核提取物将显示硒蛋白本身或硒结合蛋白是否由膳食硒摄入诱导，在 GSH-Px 的调节中发挥作用基因。