Recombination in yeast

酵母中的重组

• 批准号: 8099534
• 负责人: THOMAS PETES
• 金额: $ 45.73万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8099534
• 关键词: Affect; Alleles; Cells; Chromosome Arm; Chromosome Mapping; Chromosome Structures; Chromosomes; Coupled; Critical Pathways; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Sequence; Development; Disease; Eukaryota; Event; Frequencies; Friedreich Ataxia; Fungal Genome; Gamma Rays; Gene Conversion; Generations; Genes; Genetic; Genetic Recombination; Genome; Genomics; Goals; Human; Lead; Malignant Neoplasms; Maps; Meiosis; Meiotic Recombination; Methods; Microarray Analysis; Mismatch Repair; Mitotic; Mitotic Recombination; Mutation; Organism; Prokaryotic Cells; Property; Proteins; Regulation; Research; Resolution; Saccharomyces cerevisiae; System; Testing; Time; Trinucleotide Repeats; Tumor Suppressor Genes; Ultraviolet Rays; Yeasts; base; chromosome loss; improved; neoplastic cell; novel; repaired; research study

DESCRIPTION (provided by applicant): The ability to repair double-stranded DNA breaks (DSBs) by recombination is a universal feature of both prokaryotes and eukaryotes. In addition, mitotic recombination is an important mechanism in the generation of a tumor cell, since recombination can lead to loss of the wild-type allele of a tumor suppressor gene in a cell that is heterozygous for such a mutation. The goal of the proposed studies is to understand the mechanisms of mitotic recombination. Because of the universality of recombination and DSB repair, our experiments, which will be done with the yeast Saccharomyces cerevisiae, will improve our understanding of recombination in higher eukaryotes, including humans. The first Specific Aim is to characterize spontaneous mitotic recombination events. Because mitotic events are 104-fold less frequent than meiotic exchanges, the mechanism of mitotic recombination has been investigated much less extensively than meiotic recombination. We recently developed a novel genetic method of selecting reciprocal mitotic crossovers. We will use this method, coupled with microarray technology, to make the first high-resolution genetic map based on mitotic recombination (Specific Aim IA and ID). This analysis should identify regions of high (hotspots) and low (coldspots) exchange. We will determine whether mitotic hotspots and coldspots correlate with meiotic hotspots and coldspots (Specific Aim IB). We will also use this system to test whether certain DNA sequences stimulate crossovers (Specific Aim IC). One of the sequences that we will test (poly AAG) is associated with the human triplet-repeat-expansion disease Friedreich's ataxia. Our second Specific Aim is to investigate the regulation of spontaneous mitotic recombination events. We will examine the proteins required to catalyze reciprocal crossovers, as well as the proteins required to regulate their frequency. The third Specific Aim is to map DNA damage-induced mitotic crossovers at high resolution, both events induced by ultraviolet light and by gamma radiation. The analysis of DNA-damage induced mitotic crossovers will be done non-selectively throughout the genome, the first such analysis done in any organism.

描述（由申请人提供）：通过重组修复双链DNA断裂（DSB）的能力是原核生物和真核生物的普遍特征。此外，有丝分裂重组是肿瘤细胞生成的重要机制，因为重组可导致此类突变杂合的细胞中肿瘤抑制基因的野生型等位基因的丢失。拟议研究的目的是了解有丝分裂重组的机制。由于重组和 DSB 修复的普遍性，我们使用酿酒酵母进行的实验将提高我们对高等真核生物（包括人类）重组的理解。第一个具体目标是表征自发有丝分裂重组事件。由于有丝分裂事件的频率比减数分裂交换低 104 倍，因此对有丝分裂重组机制的研究远不如减数分裂重组广泛。我们最近开发了一种选择相互有丝分裂交叉的新遗传方法。我们将使用这种方法，结合微阵列技术，制作第一个基于有丝分裂重组的高分辨率遗传图谱（Specific Aim IA和ID）。该分析应确定高（热点）和低（冷点）交换的区域。我们将确定有丝分裂热点和冷点是否与减数分裂热点和冷点相关（具体目标 IB）。我们还将使用该系统来测试某些 DNA 序列是否会刺激交叉（Specific Aim IC）。我们将测试的序列之一（多聚 AAG）与人类三联体重复扩增疾病弗里德赖希共济失调有关。我们的第二个具体目标是研究自发有丝分裂重组事件的调节。我们将检查催化相互交叉所需的蛋白质，以及调节其频率所需的蛋白质。第三个具体目标是以高分辨率绘制 DNA 损伤诱导的有丝分裂交叉，这两种事件都是由紫外线和伽马辐射诱导的。 DNA 损伤诱导的有丝分裂交叉的分析将在整个基因组中非选择性地进行，这是在任何生物体中首次进行的此类分析。