Mutations Arising During DNA Repair

描述(申请人提供)：许多人类疾病都与等位基因有关，这些等位基因可以是简单的单核苷酸改变为拷贝数变异、基因融合和易位。最近的全基因组测序的丰富使人们意识到，最近的突变与远古出现的变异一样发挥着重要作用。这些与疾病相关的等位基因可由DNA复制过程中以多种方式引起的错误引起的突变事件引起，包括(A)电离辐射，(B)可移动遗传元件的切除，(C)在停滞的复制叉处形成双链断裂(DSB)，(D)形成替代二级结构的DNA序列中的复制滑移，(E)复制叉子和转录机制之间的冲突，以及(F)单链DNA的填充。该计划项目专注于高度互动的一系列项目中的这些不同的突变来源，这些项目将布兰代斯大学和塔夫茨大学的研究人员联系在一起，研究三种强大的模式生物：细菌磅大肠杆菌、单细胞真核生物、发芽酵母酿酒酵母和后生动物、果蝇D.黑腹果蝇。我们的目标是非常详细地了解几种重要类型突变的产生方式，并识别其缺陷会提高其外观的转录、复制和DNA修复因子。重要的是，拟议的核心设施将使我们能够将我们的研究扩展到以前可能的范围之外，并在基因组范围内测试我们的模型。通过作为本计划项目组成部分提出的核心设施，参与者将把最先进的高通量技术应用于他们的修复相关突变研究。数据收集和分析的自动化将使每个调查员能够扩大他们的调查范围，并将建立研究突变产生的新的标准程序。将通过两个机构的整个实验室每两个月举行一次会议，以及两名外部专家顾问参加的务虚会来促进互动。

DESCRIPTION (provided by applicant): Many human diseases are associated with alleles that can be as simple as single-nucleotide changes to copy-number variants, gene fusions and translocations. The wealth of recent whole-genome sequencing has led to the realization that recent mutation play as important a role as variations that arose in distant ancestors. These disease-associated alleles can arise from mutational events that are provoked by errors arising In a number of ways during DNA replication, including (a) ionizing radiation, (b) the excision of mobile genetic elements, (c) the formation of double strand breaks (DSBs) at stalled replication forks, (d) replication slippage in DNA sequences that form alternative secondary structures, (e) conflicts between replication forks and the transcription machinery, and (f) the filling-in of singe-stranded DNA. This Program Project focuses on these different sources of mutation in a highly interactive set of projects that link researchers at Brandeis University and Tufts University studying three powerful model organisms: the bacterium pound coli, the single cell eukaryote, the budding yeast S. cerevisiae, and the metazoan, fruit fly D. melanogaster. Our goal is to understand in great detail the way several important types of mutations arise and to identify transcription, replication and DNA repair factors whose defects elevate their appearance. Importantly, the proposed core facilities will enable us to expand our studies beyond what has been previously possible and to test our models on genome-wide scales. Through the Core Facilities proposed as an integral part of this Program Project, participants will apply state-of-the-art high-throughput technologies to their studies of repair-associated mutagenesis. Automation of data collection and analysis will enable each investigator to expand the scope of their Investigation and will establish new standard procedures for the study of mutation generation. Interactions will be fostered by bi-monthly meetings of the entire labs from the two institutions, as well as at a retreat attended by two outside expert advisors.