Role of regulation of eukaryotic DNA replication in preserving genome stability

真核 DNA 复制调控在保持基因组稳定性中的作用

基本信息

批准号：
9106635
负责人：
JOACHIM J LI
金额：
$ 35.66万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-07-01 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9106635
关键词：
Address Anaphase Aneuploidy Behavior monitoring Biological Biological Assay Biological Models Biology Cell Cycle Cell division Cells Cellular biology Centromere Chromatids Chromosomal Breaks Chromosomes Complex DNA DNA biosynthesis DNA replication fork Defect Dependence Ensure Eukaryota Eukaryotic Cell Event Evolution Frequencies Gene Amplification Gene Duplication Genes Genetic Recombination Genetic Variation Genome Genome Stability Genomics Goals Human Lead Link Malignant Neoplasms Mammalian Cell Measures Mitosis Mitotic Molecular Molecular Evolution Molecular Genetics Mutate Mutation Oncogenes Pathology Pathway interactions Probability Process Proteins Publishing Regulation Repetitive Sequence Replication Initiation Replication Origin Role Saccharomyces cerevisiae Saccharomycetales Sister Chromatid Source Speed System Techniques Technology Testing Thinking Work Yeasts arm cell growth cohesion daughter cell genetic analysis genetic signature homologous recombination insight interest public health relevance research study single molecule spindle pole body tool tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): A central tenet of eukaryotic cell biology is that DNA replication must be tightly controlled so that it occurs only once per cell cycle. We study replication control in budding yeast because this model system offers an exceptional opportunity to dissect the complex, overlapping mechanisms that are required to achieve this control with such extraordinary fidelity. Additionally, the molecular genetic tools available in budding yeast allow us to apply both simple and sophisticated technologies to query the effects of disrupting replication control. Our longstanding work has been instrumental in revealing multiple overlapping mechanisms used by eukaryotes to reduce the probability of re-initiation within a cell cycle. However, since eukaryotic genomes contains thousands of replication origins, it may be impossible to eliminate rare, sporadic re-initiation over the course of multiple cell divisions, and this problem can be significantly exacerbated by just a slight compromise in replication control. Thus one of our long-term goals is to discern the role of re-replication as a driver of genome plasticity and instability. Recently we have shown that loss of replication control is a powerful instigator of genetic alteration and variation in eukaryotes. We find that inappropriate re-initiation of DNA replication within a cell cycle can induce tandem intrachromosomal amplifications and aneuploidy by up to 104-106 folds. Here we propose to (1) investigate the mechanism of re-replication induced gene amplification; (2) investigate the mechanism of re- replication induced aneuploidy; (3) expand our ability to detect re-replication and genetic alterations arising from re-replication; and (4) explore whether re-replication provides a source of spontaneous segmental amplification during normal cellular growth. This work will establish re-replication as a potential new source of genetic variation and alteration in budding yeast that could ultimately impact how we think about molecular evolution and oncogenesis.

描述（由申请人提供）：真核细胞生物学的一个中心原则是 DNA 复制必须受到严格控制，以便每个细胞周期仅发生一次。我们研究芽殖酵母中的复制控制，因为该模型系统提供了剖析 DNA 复制的绝佳机会。此外，芽殖酵母中可用的分子遗传工具使我们能够应用简单和复杂的技术来探究破坏复制控制的影响。有助于揭示真核生物使用的多种重叠机制来降低细胞周期内重新启动的可能性。然而，由于真核基因组包含数千个复制起点，因此在整个过程中消除罕见的、零星的重新启动可能是不可能的。多次细胞分裂，而且复制控制中的轻微妥协就会显着加剧这个问题，因此我们的长期目标之一是识别重新复制作为基因组可塑性和不稳定性驱动因素的作用。控制是真核生物遗传改变和变异的强大刺激因素，我们发现细胞周期内DNA复制的不适当重新启动可以诱导串联染色体内扩增和非整倍性。 104-106倍。这里我们建议（1）研究再复制诱导的基因扩增的机制；（2）研究再复制诱导的非整倍性的机制；（3）扩大我们检测再复制和遗传改变的能力。 (4)探讨再复制是否在正常细胞生长过程中提供自发片段扩增的来源。这项工作将把再复制确立为遗传变异和改变的潜在新来源。芽殖酵母最终可能会影响我们对分子进化和肿瘤发生的看法。