Mutations Arising During DNA Repair

描述（由申请人提供）：许多人类疾病都与等位基因有关，这些等位基因可以简单到单核苷酸变化、拷贝数变异、基因融合和易位。最近全基因组测序的丰富成果使人们认识到，最近的突变与遥远祖先中出现的变异一样重要。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产生的错误引起的突变事件产生，包括(a)电离辐射，(b)可移动遗传元件的切除，(c)在停滞的复制叉上形成双链断裂（DSBs）， (d)形成替代二级结构的DNA序列的复制滑移，(e)复制叉与转录机制之间的冲突。(f)单链DNA的填充。该项目通过一系列高度互动的项目，将布兰代斯大学和塔夫茨大学的研究人员联系在一起，重点研究这些不同的突变来源，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、出芽酵母酿酒酵母和后生动物果蝇。我们的目标是非常详细地了解几种重要类型的突变产生的方式，并识别转录、复制和DNA修复因子，这些因子的缺陷会提高它们的外观。重要的是，拟议的核心设施将使我们能够扩展我们的研究，超越以前的可能性，并在全基因组范围内测试我们的模型。通过作为该计划项目组成部分的核心设施，参与者将应用最先进的高通量技术进行修复相关突变的研究。数据收集和分析的自动化将使每个研究者能够扩大他们的调查范围，并将为突变产生的研究建立新的标准程序。这两个机构的整个实验室的双月会议，以及由两名外部专家顾问参加的静修会，将促进相互作用。

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.