Regulation of DNA recombination in Saccharomyces cerevisiae

酿酒酵母 DNA 重组的调控

• 批准号: 7413253
• 负责人: Grzegorz A Ira
• 金额: $ 28.5万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7413253
• 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; Essential Genes; Event; Evolution; Fanconi' s Anemia; Fluorescence; Fluorescence Microscopy; Genes; Genetic; Genetic Crossing Over; Genetic Epistasis; Genetic Nondisjunction; Genetic Recombination; Genome; Genome Stability; Genomic Instability; Genomics; Goals; Green Fluorescent Proteins; 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; Outcome; Pathway interactions; Predisposition; Pregnancy; Pregnancy loss; Process; Proteins; Recruitment Activity; Regulation; Research; Research Personnel; Resolution; S cerevisiae DNA2 protein; Saccharomyces cerevisiae; Saccharomycetales; Site; Techniques; Trisomy; Two-Dimensional Gel Electrophoresis; Yeasts; chromatin immunoprecipitation; endonuclease; fluorescence microscope; gene therapy; helicase; homologous recombination; human subject; irradiation; mutant; novel strategies; prevent; programs; 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(申请人提供)：我们的长期目标是通过重组了解DMA修复过程的机制。DNA重组修复自发发生或由化学物质或辐射引起的DNA双链断裂(DSB)和缺口。根据细胞状态的不同，相同类型的DNA损伤可以通过不同的重组途径修复。决定以何种方式修复损伤对细胞维持基因组稳定至关重要。本项目的目的是了解在模式生物酿酒酵母中选择重组途径的机制。DNA修复过程在进化过程中是保守的，因此所提出的研究对于我们理解人类DNA重组修复的机制具有很高的相关性。我们使用了一种新的方法来识别参与DNA重组调控的重要蛋白质。应用荧光显微镜、染色质免疫沉淀和双向凝胶电泳法，研究重组途径选择的机制和相关蛋白的功能。我们将确定同源和非同源双链断裂修复途径之间细胞周期调节的分子开关的机制。这项研究对人类细胞基因治疗的新策略具有重要意义。我们已经开发了一种方法来鉴定和表征调控交叉和非交叉重组选择的因素。交叉途径控制对于避免有丝分裂周期中杂合性丢失、基因组重排以及防止染色体不分离导致减数分裂细胞中的非整倍体(三体或单体)至关重要。非整倍体是人类最常见的染色体异常，在所有临床确认的妊娠中至少有5%发生，是导致妊娠丢失的主要遗传原因。我们研究的蛋白质编码基因的突变会导致人类患上严重的疾病，从而增加癌症的易感性。因此，在基因和分子水平上了解它们的功能对人类健康是一个至关重要的课题。