Mutation spectra in DNA repair-deficient backgrounds

这项研究将直接解决有关DNA修复系统在塑造动物基因组中自发突变过程的速率，模式和适应性后果中的作用的基本问题。六个秀丽隐杆线虫基因，所有在酵母和人类中广泛表征的切除DNA修复基因的直向同源物，将通过UV-三甲基补骨脂素（UV-TMP）诱变缺失，并且将对六个敲除菌株中的每一个启动突变累积实验。核和线粒体基因组中的突变率和模式将通过使用直接DNA测序方法检测每组修复缺陷章动累积（MA）系中的突变来评价。研究了C. elegans将被直接评估。在一组长期的野生型C.秀丽线虫MA系将提供一个前所未有的标准，对照该标准，将用生活史特征测定来检查修复缺陷背景中的突变。关于一组长期野生型C.秀丽线虫MA系将提供一个前所未有的标准，修复缺陷MA系中的突变将与之直接比较。这项研究的执行将最终在更好地了解切除DNA修复因子在形成突变过程中的作用，这些突变过程是遗传变异，人类遗传疾病和基因组进化燃料的最终来源。

This study will directly address fundamental questions about the roles of DNA repair systems in shaping the rates, patterns and fitness consequences of spontaneous mutation processes in an animal genome. Six Caenorhabditis elegans genes, all orthologues of excision DNA repair genes extensive characterized in yeast and humans, will e deleted by UV-trimethylpsoralen (UV-TMP) mutagenesis and mutation- accumulation experiments will be initiated for each of the six knockout strains. Mutation rates and patterns in both the nuclear and mitochondrial genomes will be evaluated by detection mutations in each set of repair- deficient nutation-accumulation (MA) lines using direct DNA sequencing approaches. The spectra of mutation prevented by and extent of overlap between different repair pathways in C. elegans will be directly evaluated. The fitness consequences of spontaneous mutation in a long-term set of wild-type C. elegans MA lines will provide an unprecedented standard against which mutations in repair-deficient backgrounds will be examined with life history character assays. Background information on the rates, patterns and fitness consequences of spontaneous mutation in a long-term set of wild-type C. elegans MA lines will provide an unprecedented standard against which mutations in repair-deficient MA lines will be directly compared. Execution of this study will culminate in a greater understanding of the roles of excision DNA repair factors in shaping mutation processes that are the ultimate source of genetic variation, human genetic disorders and fuel for genome evolution.