Nuclear Structure and Metazoan Origins of Replication

核结构和后生动物复制起源

• 批准号: 9505907
• 负责人: David Gilbert
• 金额: $ 37.5万
• 依托单位: SUNY, Upstate Medical University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-15 至 1998-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9505907&HistoricalAwards=false
• 关键词: Nuclear; Structure; Metazoan; Origins; Replication

9505907 Gilbert Successful development of the first cell-free system in which DNA replication is preferentially initiated at a physiologically utilized origin of replication has been achieved by this laboratory. These initial observations will be fully investigated by using a combination of cell and molecular biological tools to identify the components of nuclear structure that designate an origin of replication. Specific aims are: 1. To determine if origins of replication are designate at the time of nuclear assembly, 2. To determine if pre-designated origins in Gl nuclei are disrupted by a Xenopus early cleavage mitotic cycle, 3. To determine if the mammalian nuclear membrane is necessary for site-specific initiation of replication in Xenopus egg extracts. %%% DNA replication is central to the life cycle of every living organism and, although much is known about the process in simple organisms, the regulatory mechanisms in higher eukaryotes are largely unknown. DNA replication initiates within defined chromosomal loci in eukaryotic cells but naked DNA in systems derived from metazoan organisms do not have any such requirements. This proposal describes a novel approach based upon the hypothesis that DNA replication in metazoan chromosomes is regulated by aspects of nuclear structure that cannot be easily reconstructed when starting with cloned DNA templates. Cell-free extracts from Xenopus eggs provide an excellent model system in which to test the hypothesis that the structural and functional organization of the nucleus is crucial in regulating DNA replication. *** ??

9505907 吉尔伯特成功地开发了第一个无细胞系统，其中DNA复制优先在生理上利用的复制起点开始。这些初步的观察结果将通过使用细胞和分子生物学工具的组合进行充分研究，以确定指定复制起点的核结构的组成部分。 具体目标是：1。为了确定复制起点是否在核组装时被指定，2.为了确定G1细胞核中预先指定的起源是否被非洲爪蟾早期卵裂有丝分裂周期破坏，3.确定哺乳动物核膜是否是非洲爪蟾卵提取物中复制位点特异性启动所必需的。 DNA复制是每个生物体生命周期的核心，尽管对简单生物体中的过程了解很多，但高等真核生物中的调节机制在很大程度上是未知的。 在真核细胞中，DNA复制起始于确定的染色体位点内，但在来自后生动物生物体的系统中的裸DNA没有任何这样的要求。该提案描述了一种新的方法，该方法基于这样的假设，即后生动物染色体中的DNA复制受核结构方面的调节，当从克隆的DNA模板开始时，核结构方面不能容易地重建。非洲爪蟾卵的无细胞提取物提供了一个很好的模型系统，在其中测试的假设，即细胞核的结构和功能组织是至关重要的调节DNA复制。 *** ??