MOLECULAR GENETICS OF MAMMALIAN CYTOCHROME C OXIDASE

哺乳动物细胞色素 C 氧化酶的分子遗传学

• 批准号: 6179776
• 负责人: LAWRENCE GROSSMAN
• 金额: $ 18.71万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-02-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6179776
• 关键词: DNA binding protein; DNA footprinting; cell differentiation; cell line; cytochrome oxidase; enzyme induction /repression; enzyme mechanism; enzyme structure; gene deletion mutation; gene expression; gene interaction; genetic promoter element; genetic regulatory element; immunocytochemistry; in situ hybridization; isozymes; luciferin monooxygenase; mitochondrial membrane; protein isoforms; site directed mutagenesis; transcription factor; transfection; yeast two hybrid system

DESCRIPTION: Cytochrome c oxidase, the terminal protein of the electron transport chain, is composed in mammals of thirteen subunits, three encoded by mitochondrial DNA and ten by nuclear DNA. At least three of these nuclear subunits occur in more than one molecular form (isoforms), the products of separate genes. The proposed experiments will provide insights into the cellular function of these isoforms, which may be related to both ontogenetic development and cellular physiology. In addition, isoform expression has been implicated in human neuromuscular disease. This project will focus on three genes, already isolated, that encode two subunits, COX VIIa and COX VIIc. Subunit VIIa displays a heart/muscle-specific isoform (H) and an isoform (L) expressed in all tissue. A second subunit, VIIc, appears to have a single expressed form in all tissues. The specific aims are to (1) characterize the core promoter interactions in the COX7AL, COX7AH, and COX7C genes; (2) determine the roles of promoter-distal sequences and factors in regulatory events that occur at the COX7AL, COX7AH, and COX7C core promoters by (a) identifying new regulatory sites that are core promoter distal, (b) identifying new regulatory sites and factors by application of yeast one-and two-hybrid technologies, and (c) determining the effect of new regulatory factor binding on events at the core promoter, and also the effect of distal DNA sequences per se; and (3) elucidating promoter function in cell development and differentiation. This project fits into the longer term objectives of deducing the role of the nuclear-encoded subunits and the mechanisms used to regulated oxidative function, and applying this knowledge to the study of nuclear genes in mitochondrial disease.

描述：细胞色素c氧化酶，电子的末端蛋白 运输链，由哺乳动物的13个亚基、3个亚基组成 由线粒体DNA编码，由核DNA编码。至少有三个 这些核亚基以一种以上的分子形式(异构体)存在， 不同基因的产物。拟议的实验将提供 对这些异构体的细胞功能的洞察，这可能是 与个体发育和细胞生理学有关。在……里面 此外，人类神经肌肉的异构体表达也被发现。 疾病。这个项目将专注于三个基因，已经分离出来， 编码COX VIIa和COX VIc两个亚基。亚基VIIa显示一个 心脏/肌肉特异性亚型(H)和一个亚型(L)在ALL中表达 组织。第二个亚基VIIC似乎有一个单一的表达形式 所有的纸巾。具体目标是：(1)确定核心启动子的特征 COX7AL、COX7AH和COX7C基因的相互作用；(2)决定 启动子-末端序列和因子在调控事件中的作用 发生在COX7AL、COX7AH和COX7C核心启动子上，通过(A)识别 作为核心启动子末端的新调控位点，(B)识别新的 酵母单杂交和双杂交应用的调控位点和调控因子 (C)确定新的监管因素的影响 与核心启动子上的事件的结合以及远端DNA的影响 序列本身；(3)阐明启动子在细胞中的功能 发展与差异化。这个项目符合较长期的要求。 推断核编码亚基的作用的目标和 用于调节氧化功能的机制，并应用这一点 对线粒体疾病核基因研究的认识。