Protein: DNA Interactions in Drosophila Mitchondrial Regulation

9600681卡古尼拟议工作的长期目标是了解线粒体染色体的结构及其表达和复制对线粒体功能的调节。高等动物细胞需要线粒体，线粒体的生物发生需要线粒体DNA(MtDNA)基因组的表达和复制。由于转录和复制都受到DNA模板结构的影响，因此在体内维持DNA构象的核蛋白复合体在这些功能上的作用值得详细研究。为了阐明与线粒体染色体相关的蛋白质在维持其结构中的作用并研究它们对其功能的影响，我们将从黑腹果蝇中分离线粒体类核，纯化并鉴定其蛋白质组分，并开始克隆它们。我们的长期目标将是开发利用线粒体酶组分和核蛋白复合体形式的线粒体DNA的果蝇线粒体体外转录和复制系统。线粒体类核糖体的分离和鉴定是建立果蝇线粒体DNA功能模型的重要组成部分。线粒体是细胞中的能量产生单位，是生命所必需的。由于线粒体染色体的大小有限，线粒体生物发生所需的所有蛋白质都编码在细胞核内的线粒体之外。这些蛋白质在动物细胞中相对罕见，因此很难研究。果蝇是一种高等动物，已被证明是通过生化、遗传和发育方法研究细胞功能的优秀模型。线粒体DNA的结构和组织在动物中是高度保守的，目前的数据表明线粒体DNA包装和基因表达的机制也将是高度保守的。我们的长期目标是发展果蝇作为研究这些过程的动物模型系统。线粒体染色体及其蛋白质组分的分离和鉴定是建立果蝇动物线粒体DNA功能模型的第一步。***

9600681 Kaguni The long term objective of the proposed work is to understand the regulation of mitochondrial function, in terms of the structure of the mitochondrial chromosome and its expression and replication. Mitochondria are required in higher animal cells, and mitochondrial biogenesis requires expression and duplication of the mitochondrial DNA (mtDNA) genome. Because both transcription and replication are influenced by the structure of the DNA template, the role of nucleoprotein complexes that are involved in maintaining DNA conformation in vivo on these functions warrants detailed study. To begin to elucidate the role of proteins associated with the mitochondrial chromosome in maintaining its structure and to examine their influence on its function, we will isolate the mitochondrial nucleoid from Drosophila melanogaster, purify and characterize its protein components, and initiate efforts to clone them. Our long-term goal will be to develop in vitro transcription and replication systems from Drosophila mitochondria that employ mitochondrial enzyme fractions and mitochondrial DNA in the form of nucleoprotein complexes. The isolation and characterization of the mitochondrial nucleoid is an integral part in the development of the Drosophila model for animal mtDNA function. %%% Mitochondria are the energy-producing units in the cell and are essential to life. Because of the limited size of the mitochondrial chromosome, all of the proteins required for mitochondrial biogenesis are encoded outside the mitochondrion in the nucleus. These proteins are comparatively rare in the animal cell, and are therefore difficult to study. The fruit fly Drosophila is a higher animal that has proven to be an excellent model for the study of cellular function by biochemical, genetic and developmental approaches. The structure and organization of mitochondrial DNA (mtDNA) is highly conserved in animals, and current data suggest that the mechanisms of mtDNA packaging and gene expression will also be. Our lo ng-term goal is to develop Drosophila as an animal model system to study these processes. The isolation and characterization of the mitochondrial chromosome and its protein components represents the first step in the development of the Drosophila model for animal mtDNA function. ***