CHROMOSOME ARCHITECTURE

染色体结构

• 批准号: 2189905
• 负责人: Marc R. Gartenberg
• 金额: $ 15.92万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-12 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2189905
• 关键词: CHROMOSOME; ARCHITECTURE

The long-range goal of this project is to understand parameters which determine spatial and functional organization of DNA in the nucleus. Specifically, the work will identify chromosomal DNA sites which are anchored to immobile objects in yeast Saccharomyces cerevisiae. DNA immobilization by specific DNA sequences may represent a fundamental unit of chromosome architecture; DNA anchors may segregate chromosomal DNA into independent structural domains for DNA packaging, regulated gene expression and positioning within the nucleus. DNA anchoring sequences and associated proteins will be characterized using a newly developed molecular biological technique. The approach employs site specific recombination in vivo to generate well-defined DNA rings which contain a single promoter and a DNA fragment to be tested. Transcription of anchored DNA rings ina yeast topoisomerase mutants leads to diagnostic changes in DNA topology, in accordance with the twin domain model of transcriptional supercoiling. models of DNA organization based on analyses of biochemically manipulated chromosomes, termed nuclear scaffold, argue that DNA is arranged into loops with specific DNA segments anchoring the bases of the loops to an organizational framework. The excision methodology will be used to investigate whether sequences which bind yeast scaffold preparations function as DNA anchors in vivo. Scaffold attachment regions (SARs) to be examined include chromosomal components such as telomeres, silencers, centromeres and origins of replication. DNA anchor formation. The cell- cycle dependent anchoring of centromeres and replication origins will be determined in synchronized cell cultures. A selection system will be developed to isolate sequences with DNA anchoring activity from libraries of the sequenced yeast Chromosome III. The DNA loop structure of specific chromosomal regions will be determined by mapping the DNA anchoring elements which reside within the regions. The influence of genomic DNA anchors on plasmid partitioning and gene expression will be examined.

该项目的远程目标是了解参数 确定核中DNA的空间和功能组织。 具体而言，该工作将确定染色体DNA位点 锚定在酿酒酵母中的酵母菌中的固定物体上。 脱氧核糖核酸 特定DNA序列的固定化可能代表一个基本单位 染色体建筑； DNA锚可能将染色体DNA隔离到 DNA包装，调控基因表达的独立结构结构域 并定位在细胞核内。 DNA锚定序列和相关 蛋白质将使用新开发的分子生物学来表征 技术。 该方法在体内采用特定地点重组 生成定义明确的DNA环，其中包含单个启动子和DNA 片段要测试。 锚定DNA环的转录 拓扑异构酶突变体导致DNA拓扑的诊断变化， 根据转录超螺旋的双域模型。 基于生化操纵的分析DNA组织的模型 染色体称为核支架，认为DNA被排列成环 用特定的DNA片段将环的基础锚定为 组织框架。 切除方法将用于 研究结合酵母支架制剂的序列是否 在体内充当DNA锚固。 脚手架依恋区（SARS）为 所检查的包括染色体成分，例如端粒，消音器， 复制的中心和起源。 DNA锚形成。 细胞 - 中心粒和复制起源的循环依赖性锚定为 在同步的细胞培养物中确定。 选择系统将是 开发用于与库中DNA锚定活性分离的序列 测序的酵母染色体III。 特定的DNA环结构 染色体区域将通过映射DNA锚定确定 位于区域内的元素。 基因组DNA的影响 将检查对质粒分配和基因表达的锚固。