Genome stability in rad mutants

rad 突变体的基因组稳定性

• 批准号: 6887432
• 负责人: ADAM M BAILIS
• 金额: $ 32.38万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6887432
• 关键词: DNA damage; DNA repair; Saccharomyces cerevisiae; chromosome translocation; fungal genetics; gene deletion mutation; gene mutation; gene rearrangement; genetic recombination; immunoprecipitation; intermolecular interaction; nucleic acid repetitive sequence; polymerase chain reaction; radiation dosage; radiation genetics; radiation sensitivity; radiobiology; southern blotting

DESCRIPTION (provided by applicant): Homologous recombination is an important pathway for the repair of DNA damage by radiation, chemicals, and endogenous cellular processes. However, recombination must be carefully controlled as deleterious genome rearrangements can result from unrestricted recombination between numerous, dispersed repetitive sequences. Since the majority of these repeats are 300 bp or shorter in eukaryotes, the control of recombination between these short sequences is more stringent than for longer sequences. Several of the genes known to mediate the response to radiation in the budding yeast S. cerevisiae also control short-sequence recombination (SSR). The goal of the proposed research program is to further investigate important interactions between several of the most critical factors at the genetic and molecular levels. DNA fragment insertion assays in cells with mutant SSR factors will be used to explore the effect of specific defects, both alone and in combination, on SSR. Repair of a genomic double-strand break by recombination will be used to monitor the kinetics of SSR in these mutants. Ligation mediated PCR will be employed to track and quantitate secondary breaks created during SSR. Chromatin immunoprecipitation will be used to follow the interaction of several important protein complexes and the recombination substrates. This combination of genetic and physical methods will promote a more thorough understanding of several of the molecular interactions critical for SSR, and the maintenance of genome stability.

描述（由申请人提供）：同源重组是修复辐射、化学物质和内源性细胞过程造成的 DNA 损伤的重要途径。 然而，重组必须小心控制，因为大量分散的重复序列之间的无限制重组可能导致有害的基因组重排。 由于真核生物中大多数重复序列的长度为 300 bp 或更短，因此对这些短序列之间重组的控制比对较长序列的重组控制更为严格。 已知在芽殖酵母酿酒酵母中介导辐射反应的几个基因也控制短序列重组（SSR）。 拟议研究计划的目标是进一步研究遗传和分子水平上几个最关键因素之间的重要相互作用。 具有突变 SSR 因子的细胞中的 DNA 片段插入测定将用于探索特定缺陷（单独或组合）对 SSR 的影响。 通过重组修复基因组双链断裂将用于监测这些突变体中 SSR 的动力学。 连接介导的 PCR 将用于跟踪和定量 SSR 期间产生的二次断裂。 染色质免疫沉淀将用于跟踪几种重要的蛋白质复合物和重组底物的相互作用。 这种遗传和物理方法的结合将促进对 SSR 和维持基因组稳定性至关重要的几种分子相互作用的更深入理解。