Single Stranded DNA: The Genome's Achilles Heel

单链 DNA：基因组的致命弱点

基本信息

批准号：
8392278
负责人：
WENYI FENG
金额：
$ 21.28万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-30 至 2014-11-30
项目状态：
已结题

项目摘要

DNA replication is a fundamental process on which all organisms depend for faithful passage of genetic material from one generation to the next. The DNA replication process is often "stressed" either by mutations or by exogenous reagents that can cause DNA damage or impediments to the replication machinery. Replication stress can give rise to genomic instability and may ultimately lead to diseases including cancer. My main research interest is how defective replication cause genomic instability. The general hypothesis governing this proposal is that single-stranded DNA (ssDNA) production upon replication fork stalling and destablization in replication checkpoint-deficient mutants is a potentially lethal event that prevents complete synthesis of the genome, causes chromosomal breakage, and ultimately genomic instability. The specific aims of this project are: 1. Investigate the link between extensive ssDNA formation during replication stress and chromosome breakage using the genome-wide ssDNA and chromosome breakage mapping techniques developed by the applicant in combination with other genetic approaches. 2. Elucidate the mechanism/cause of replication fork instability in checkpoint mutants in HU by examining the structural/compositional changes that occur in the replication complex upon replication stress by a minichromosome purification system coupled with mass spectrometry. 3. Identify origins of replication and chromosome fragile sites in humans using a combination of ssDNA and chromosome breakage mapping that have been adapted for the mammalian systems.

DNA复制是一个基本过程，所有生物都依赖于遗传的忠实通过从一代到另一代的材料。 DNA复制过程通常是由突变或通过外源试剂可能造成DNA损伤或障碍的复制机械。复制应力会导致基因组不稳定，并最终导致疾病包括癌症。我的主要研究兴趣是，有缺陷的复制如何导致基因组不稳定性。这统治此提案的一般假设是复制后单链DNA（ssDNA）产生复制检查点缺陷突变体中的叉子失速和破坏性是一个潜在的致命事件防止基因组的完全合成，导致染色体断裂，并最终导致基因组不稳定。该项目的具体目的是： 1。研究复制应力和染色体过程中广泛的ssDNA形成之间的联系使用全基因组的ssDNA和染色体破裂图映射技术的破裂申请人与其他遗传方法结合使用。 2。通过检查检查点突变体中复制叉不稳定性的机制/原因在复制络合物中发生的结构/成分变化在复制应力后通过微小浓粒子体发生纯化系统与质谱法结合。 3。使用ssDNA和已适用于哺乳动物系统的染色体断裂映射。