Regulation of DNA recombination in Saccharomyces cerevisiae

酿酒酵母 DNA 重组的调控

• 批准号: 7896066
• 负责人: Grzegorz A Ira
• 金额: $ 27.6万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-17 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7896066
• 关键词: Aneuploidy; Animal Model; Biological Assay; Bloom Syndrome; Cell Cycle; Cell Cycle Stage; Cells; Chemicals; Chimeric Proteins; Chromosome Segregation; Chromosome abnormality; Chromosomes; Collection; Cruciform DNA; Cyclin-Dependent Kinases; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Sequence Rearrangement; Development; Disease; Double Strand Break Repair; Enzymes; Event; Evolution; Fanconi' s Anemia; Fluorescence Microscopy; Genes; Genetic; Genetic Crossing Over; Genetic Epistasis; Genetic Nondisjunction; Genetic Recombination; Genome; Genome Stability; Genomic Instability; Genomics; Goals; Health; Holliday Junction Resolvases; Homologous Gene; Human; Invaded; Length; Loss of Heterozygosity; Malignant Neoplasms; Meiosis; Meiotic Recombination; Mitotic; Mitotic Recombination; Molecular; Molecular Genetics; Monosomy; Mutation; Orthologous Gene; Outcome; Pathway interactions; Predisposition; Pregnancy; Pregnancy loss; Process; Proteins; Proteome; Recruitment Activity; Regulation; Research; Resolution; Role; S cerevisiae DNA2 protein; Saccharomyces cerevisiae; Saccharomycetales; Site; Techniques; Trisomy; Two-Dimensional Gel Electrophoresis; Yeasts; chromatin immunoprecipitation; endonuclease; gene therapy; helicase; homologous recombination; human subject; irradiation; mutant; novel strategies; prevent; protein function; recombinational repair; repaired; research study

DESCRIPTION (provided by applicant): Our long-term goal is to understand the mechanisms of DMA repair process via recombination. DNA recombination repairs DNA double-strand breaks (DSBs) and gaps that occur spontaneously or are induced by chemicals or irradiation. The same type of DNA damage can be repaired by different pathways of recombination, depending on the cellular state. The decision as to which way to repair damage is crucial for cells to maintain genome stability. The objective of this project is to understand the mechanisms underlying the choice of recombination pathway in the model organism Saccharomyces cerevisiae. DNA repair processes are conserved in evolution, so the proposed research is highly relevant to our understanding of the mechanisms of DNA repair via recombination in humans. We used a new approach to identify important proteins involved in DNA recombination regulation. Fluorescence microscopy, chromatin immunoprecipitation and two-dimensional gel electrophoresis, we applied to study the mechanisms of recombination pathway choice and the function of proteins involved. We will define the mechanism of the cell cycle-regulated molecular switch between homologous and nonhomologous DSBs repair pathways. This study has important implications for new strategies for gene therapy in human cells. We have developed an assay to identify and characterize factors regulating choice of crossover and noncrossover recombination. Crossover pathway control is crucial to avoid loss of heterozygosity, genomic rearrangements in mitotic cycle and to prevent chromosome nondisjunction that causes aneuploidy (trisomy or monosomy) in meiotic cells. Aneuploidy is the most commonly identified chromosome abnormality in humans, occurring in at least 5% of all clinically recognized pregnancies and is the leading genetic cause of pregnancy loss. Mutations in genes encoding the proteins we study cause severe disorders in humans with increased predisposition to cancer. Therefore understanding their function, on the genetic and molecular levels is a critically important subject for human health.

描述（由申请人提供）：我们的长期目标是了解通过重组进行 DMA 修复过程的机制。 DNA 重组可修复自发发生或由化学物质或辐射诱导的 DNA 双链断裂 (DSB) 和间隙。根据细胞状态，相同类型的 DNA 损伤可以通过不同的重组途径进行修复。选择哪种方式修复损伤对于细胞维持基因组稳定性至关重要。该项目的目的是了解模式生物酿酒酵母中重组途径选择的潜在机制。 DNA 修复过程在进化中是保守的，因此拟议的研究与我们对人类通过重组进行 DNA 修复机制的理解高度相关。我们使用了一种新方法来鉴定参与 DNA 重组调控的重要蛋白质。我们应用荧光显微镜、染色质免疫沉淀和二维凝胶电泳来研究重组途径选择的机制和相关蛋白的功能。我们将定义同源和非同源 DSB 修复途径之间细胞周期调节分子转换的机制。这项研究对于人类细胞基因治疗的新策略具有重要意义。我们开发了一种测定法来识别和表征调节交叉和非交叉重组选择的因素。交叉途径控制对于避免杂合性丧失、有丝分裂周期中的基因组重排以及防止减数分裂细胞中导致非整倍性（三体性或单体性）的染色体不分离至关重要。非整倍体是人类最常见的染色体异常，在所有临床认可的妊娠中至少有 5% 发生，是导致妊娠失败的主要原因。我们研究的蛋白质编码基因突变会导致人类严重疾病，增加患癌症的可能性。因此，在遗传和分子水平上了解它们的功能对于人类健康至关重要。