Tissue Specific Expression of Cytochrome c Oxidase

细胞色素 c 氧化酶的组织特异性表达

• 批准号: 9405363
• 负责人: Margaret Lomax
• 金额: $ 20万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-15 至 1997-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9405363&HistoricalAwards=false
• 关键词: Tissue; Specific; Expression; Cytochrome; Oxidase

Abstract 9405363 The enzyme complexes of the mitochondrial electron transport chain are unique. Each complex is composed of subunits encoded in two different genomes, the mitochondrial and the nuclear genomes. Cytochrome c oxidase (COX), the last enzyme complex in the chain provides an ideal model system in which to analyze both coordinate and tissue-specific transcriptional regulation of these essential respiratory genes. The long-term goal of this project is to understand the genetic regulatory mechanisms controlling both tissue-specific expression and developmental regulation of mammalian genes for the mitochondrial-encoded COX subunits. We will (1) define skeletal muscle-specific enhancer elements in the gene for COX subunit VIII-H by means of gene fusions with either the CAT or lacZ reporter genes (2) map the sites of DNA-protein interactions between nuclear proteins and tissue-specific enhancers in the COX8H gene by DNAse hypersensitivity, DNAse footprinting and electrophoretic mobility shift assays and determine the functional significance of these interactions by gene transactivation studies (3) physically map the bovine genes for the tissue-specific isoforms of COX subunit VIII and (4) nap the COX VIII isoform genes in the mouse by inter-specific hybrid back-crosses. %%% These studies will constitute the first comprehensive analysis of the transcriptional regulation of a class of genes, the tissue- specific respiratory genes, essential for respiratory function in contractile muscle. The results should provide insight into the genetic regulation of tissue-specific subunits of an essential multi-subunit enzyme complex involved in oxidative metabolism and provide tools to enable us to address the function of these isoforms in future studies. ***

摘要9405363线粒体电子传递链的酶复合物是独特的。每个复合体由两个不同基因组（线粒体和核基因组）中编码的亚基组成。 细胞色素c氧化酶（考克斯），在链中的最后一个酶复合物提供了一个理想的模型系统，在其中分析协调和组织特异性转录调控的这些必要的呼吸基因。本项目的长期目标是了解控制哺乳动物基因的组织特异性表达和发育调节的基因调控机制，为哺乳动物编码的考克斯亚基。我们将（1）通过与CAT或lacZ报告基因的基因融合来定义考克斯亚基VIII-H基因中的骨骼肌特异性增强子元件（2）通过DNA酶超敏性来定位COX 8 H基因中核蛋白和组织特异性增强子之间的DNA-蛋白相互作用位点，DNA酶足迹法和电泳迁移率变化分析，并通过基因反式激活研究确定这些相互作用的功能意义（3）物理定位组织的牛基因-考克斯亚基VIII的特异性同种型和（4）通过种间杂交回交在小鼠中敲除考克斯VIII同种型基因。 这些研究将构成对一类基因转录调控的第一次全面分析，这类基因是组织特异性呼吸基因，对收缩肌的呼吸功能至关重要。 这些结果应该提供深入了解参与氧化代谢的必需多亚基酶复合物的组织特异性亚基的遗传调控，并提供工具，使我们能够在未来的研究中解决这些亚型的功能。 ***