Genomic studies of chimeric mitochondria in cybrids from Solanaceae

茄科杂种嵌合线粒体的基因组研究

基本信息

批准号：
7911843
负责人：
JEFFREY D PALMER
金额：
$ 5.38万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-10 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7911843
关键词：
Address Alleles Angiosperms Animals Antibiotics Applications Grants Argentina Biochemical Biological Models Biological Products Cell Nucleus Chloroplasts Collaborations DNA DNA Sequence DNA Sequence Rearrangement Data Deoxyribonuclease I Development Disease Double Strand Break Repair Elements Eukaryota Eukaryotic Cell Event Evolution Experimental Genetics Experimental Models Family Follow-Up Studies Frequencies Fruit Gene Duplication Gene Targeting Gene Transfer Genes Genetic Genetic Engineering Genetic Materials Genetic Recombination Genome Genomics Goals Grant Health Homing Homologous Gene Homologous Protein Horizontal Disease Transmission Horizontal Gene Transfer Human Hybrids Inherited Introns Knowledge Laboratory Organism Lead Length Maintenance Measures Mitochondria Mitochondrial DNA Mitochondrial Proteins Molecular Nature Nuclear Nucleotides Open Reading Frames Organism Outcome Parents Partner in relationship Pathway interactions Pattern Phylogenetic Analysis Plants Process Prokaryotic Cells Property Protoplasts Publishing RNA Reading Frames Recombinants Reporting Research Resolution Role Sampling Sequence Analysis Shapes Site Solanaceae Surveys System Testing Time Tobacco Vaccine Production Virginia Work chimeric gene driving force endonuclease gene therapy genome sequencing knowledge base microbial mitochondrial genome novel parent grant plant fungi plant genetics public health relevance rRNA Genes research study sample fixation therapeutic protein uptake

项目摘要

DESCRIPTION (provided by applicant): "Genomic studies of chimeric mitochondria in cybrids from Solanaceae" This research will be conducted primarily in Argentina at Universidad Nacional de Cuyo in collaboration with Dr. Marma Virginia Sanchez Puerta, as an extension of R01 GM070612-4. The study of mitochondrial genome evolution, dynamics, uptake of foreign DNA, and interactions with the nuclear genome is essential to a deep understanding of the eukaryotic cell. Mitochondrial genomes provide an excellent system to increase our knowledge of genome rearrangements, chimeric genes, nuclear- cytoplasmic incompatibilities, and horizontal gene transfer (HGT). Plant mitochondria are unique in their propensity to acquire genes by HGT. The most pervasive example of HGT in eukaryotes involves the cox1 intron, which encodes a putative homing endonuclease that increases the frequency of the intron's fixation via horizontal transfer. We propose to study aspects of the interactions between two distinct mitochondrial genomes that recombine in a cybrid plant obtained by protoplast fusion experiments. During cybrid formation, the two mitochondrial parental types and their genomes (only one containing the cox1 intron) will fuse, resulting in a hybrid mitochondrial genome. We will establish some 20 cybrid lines derived from somatic crosses between cox1 intron-containing and -lacking species in order to address the following two aims: First, we will test the hypothesis that the cox1 intron encodes a functional homing endonuclease in plants, assess rates of intron colonization, and measure lengths of exonic coconversion tracts that accompany intron insertion. Second, we will sequence and analyze the entire mitochondrial genomes of up to 20 cybrid lines. This will serve as an experimental model for recapitulating the natural process of HGT in plant mitochondria and will provide a rich picture of the process and pattern of mitochondrial genome recombination at various levels. Maintenance of an intact, functioning mitochondrial genome is critical for survival. Given the many molecular parallels between plants and animals, this project will be valuable for the development of both animal gene therapy and plant genetic engineering. PUBLIC HEALTH RELEVANCE: Genome rearrangements, nucleotide substitutions, and the introduction of foreign DNA shape the mitochondrial genomes of eukaryotes and often have severe consequences for human health, as evidenced by several important mitochondrially-inherited diseases. In addition, the key experimental organism of this study (tobacco) is widely used as an expression system for biopharmaceutical production of vaccines, antibiotics, and a number of therapeutic proteins. Understanding the molecular mechanisms of mitochondrial DNA evolution is therefore of considerable importance to human health and disease.

描述（由申请人提供）：“茄科杂种嵌合线粒体的基因组研究” 这项研究将主要在阿根廷库约国立大学与 Marma Virginia Sanchez Puerta 博士合作进行，作为 R01 GM070612-4 的延伸。线粒体基因组进化、动力学、外源 DNA 的摄取以及与核基因组相互作用的研究对于深入了解真核细胞至关重要。线粒体基因组提供了一个优秀的系统来增加我们对基因组重排、嵌合基因、核质不相容性和水平基因转移（HGT）的了解。植物线粒体通过 HGT 获取基因的倾向是独特的。真核生物中 HGT 最普遍的例子涉及 cox1 内含子，它编码一种假定的归巢核酸内切酶，通过水平转移增加内含子固定的频率。我们建议研究两个不同线粒体基因组之间相互作用的各个方面，这两个线粒体基因组在通过原生质体融合实验获得的细胞杂种植物中重组。在细胞杂种形成过程中，两种线粒体亲本类型及其基因组（只有一种含有 cox1 内含子）将融合，从而产生杂合线粒体基因组。我们将建立约 20 个由含有 cox1 内含子和缺乏 cox1 内含子的物种之间的体细胞杂交衍生的细胞杂种系，以实现以下两个目标：首先，我们将测试 cox1 内含子在植物中编码功能性归巢核酸内切酶的假设，评估比率内含子定植，并测量伴随内含子插入的外显子共转换束的长度。其次，我们将对多达 20 个细胞系的整个线粒体基因组进行测序和分析。这将作为一个实验模型来重现植物线粒体中 HGT 的自然过程，并将提供线粒体基因组重组在各个水平上的过程和模式的丰富图片。维持完整、功能正常的线粒体基因组对于生存至关重要。鉴于植物和动物之间有许多分子相似之处，该项目对于动物基因治疗和植物基因工程的发展都很有价值。公共卫生相关性：基因组重排、核苷酸取代和外源 DNA 的引入塑造了真核生物的线粒体基因组，并且常常对人类健康产生严重后果，几种重要的线粒体遗传性疾病就证明了这一点。此外，本研究的关键实验生物体（烟草）被广泛用作疫苗、抗生素和许多治疗性蛋白质的生物制药生产的表达系统。因此，了解线粒体 DNA 进化的分子机制对于人类健康和疾病具有相当重要的意义。