MITOCHONDRIAL DNA POLYMERASE: MECHANISM AND STRUCTURE

线粒体 DNA 聚合酶：机制和结构

• 批准号: 6476519
• 负责人: LAURIE SIMON KAGUNI
• 金额: $ 26.73万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6476519
• 关键词: DNA binding protein; DNA directed DNA polymerase; DNA footprinting; DNA replication; cell capsule; enzyme mechanism; enzyme reconstitution; enzyme structure; enzyme substrate; gel mobility shift assay; immunoprecipitation; laboratory rabbit; membrane biogenesis; mitochondria; mitochondrial DNA; mitochondrial membrane; polymerization; protein structure function; site directed mutagenesis

DESCRIPTION (adapted from applicant's abstract): The long-term objective of the proposed work is the elucidation of the mechanism of DNA replication in animal mitochondria, and its relationship to mitochondrial mutagenesis and human disease. A combined approach of current methods in biochemistry and molecular genetics is being pursued to study the mechanism, structure and physiology of the major replicative enzyme in mitochondria, DNA polymerase gamma (pol gamma), and the specific roles of the essential replication accessory protein, mitochondrial single-stranded DNA-binding protein. Structure-function studies of pol gamma will involve comparative analyses of native and altered forms, to elucidate the role of the small subunit in enzyme function and to dissect functional domains in both the catalytic and accessory subunits. Physiological studies will focus on expanding the development of a transgenic fly model to examine the relationship between pol gamma function, mitochondrial DNA replication fidelity, aging and disease. The control of animal cell reproduction during normal development, and the loss of control during cancerous development, is of central importance in the processes of human growth, aging, and disease. Mitochondrial biogenesis proceeds in parallel with cell proliferation, but it is neither tightly coupled to mitochondrial DNA replication nor to the cell cycle. Nevertheless, because both the DNA content of the mitochondrion and the number of mitochondria in cells remain relatively constant, specific regulatory mechanisms are likely required to couple mitochondrial DNA replication and biogenesis to nuclear DNA replication and cell division. A detailed analysis of the key enzyme involved in mitochondrial DNA replication, DNA polymerase gamma, will represent a major contribution toward an eventual understanding of mitochondrial biogenesis and function in normal and diseased tissues. A variety of mitochondrial DNA diseases have recently been documented. This, and an increased recognition that antiviral and antitumor drugs frequently affect mitochondrial DNA function, and in particular, the activity of po1 gamma, demonstrate a critical need for an in-depth understanding of this essential cellular DNA polymerase.

描述（改编自申请人的摘要）： 拟开展的工作是阐明动物体内DNA复制的机制 线粒体及其与线粒体突变和人类 疾病生物化学和分子生物学方法的结合 遗传学正在进行研究的机制，结构和生理， 线粒体中的主要复制酶，DNA聚合酶γ（pol γ）， 以及必需复制辅助蛋白的特定作用， 线粒体单链DNA结合蛋白。结构-功能研究 将涉及对天然和改变形式的比较分析， 阐明小亚基在酶功能中的作用， 催化亚基和辅助亚基中的功能结构域。生理 研究将集中在扩大转基因苍蝇模型的发展， 检测聚合酶γ功能、线粒体DNA 复制保真度、老化和疾病。 在正常发育过程中对动物细胞繁殖的控制， 在癌症发展过程中的控制，是至关重要的， 人类的生长、衰老和疾病的过程。线粒体生物发生 与细胞增殖平行进行，但既不是紧密耦合的 对线粒体DNA复制和细胞周期都没有影响。然而由于 线粒体的DNA含量和线粒体的数量都比对照组高。 细胞保持相对恒定，可能有特定的调节机制 将线粒体DNA复制和生物发生与核DNA结合起来 复制和细胞分裂。对所涉及的关键酶的详细分析 在线粒体DNA复制中，DNA聚合酶γ将代表一个主要的 对线粒体生物发生的最终理解的贡献， 在正常和病变组织中发挥作用。 最近已经记录了各种线粒体DNA疾病。这个， 人们越来越认识到，抗病毒和抗肿瘤药物经常 影响线粒体DNA功能，特别是po 1的活性 伽马，表明迫切需要深入了解这一点 必需的细胞DNA聚合酶