CAREER: Recovery of Replication Following DNA Damage in E.coli

在DNA损伤存在的情况下不准确的复制是在异常分裂的细胞中观察到的大多数细胞重排和突变的原因。DNA损伤，如紫外线辐射引起的损伤，严重损害了复制基因组模板的能力，并导致复制机制的停止和新复制的DNA的缺口。这项研究的总体目标是确定复制过程中遇到的损伤在体内处理和修复的细胞机制(S)。该项目利用两种先前建立的细胞分析方法来监测在体内被DNA损伤阻止的复制叉处发生的分子事件。这种方法将确定决定DNA修复或跨损伤合成何时在受阻复制叉处发挥作用以恢复DNA合成的遗传要求和条件。第二种方法将描述在紫外线照射细胞新复制的DNA中观察到的缝隙底物中处理和修复损伤的基因和机制。通过理解在DNA损伤存在的情况下如何保持忠实的复制，将阐明可能导致突变、基因组重排或细胞致死的条件和事件。此外，这个项目将通过加强本科课程，为学生创建一个新的分子遗传学实验室，并增加学生对这所大学的研究参与的过程来完成。该项目将大大增加参与研究的潜在青年科学家的数量，并提高该国这一地区的学生接触到的科学的质量。

Inaccurate replication in the presence of DNA damage is responsible for the majority of cellular rearrangements and mutagenesis that are observed in abnormally dividing cells. DNA damage, such as that induced by UV irradiation, severely impairs the ability of replication to copy the genomic template and leads to the arrest of the replication machinery and gaps in the newly replicated DNA. The overall objective of this research is to identify the cellular mechanism(s) by which lesions encountered during replication are processed and repaired in vivo. This project utilizes two previously established cellular assays to monitor the molecular events that occur at replication forks arrested by DNA damage in vivo. This approach will identify the genetic requirements and conditions that determine when DNA repair or translesion synthesis functions at arrested replication forks to restore DNA synthesis. A second approach will characterize the genes and mechanisms that operate to process and repair lesions in the gapped substrates that are observed to arise in the newly replicated DNA of UV-irradiated cells. Through an understanding of how faithful replication is maintained in the presence of DNA damage, the conditions and events that can lead to mutagenesis, genomic rearrangements, or cell lethality will be elucidated. In addition, this project will be accomplished through a process that strengthens the undergraduate curriculum, creates a new molecular genetics laboratory for students, and increases student participation in research at this university. This project will significantly increase the number of potential young scientists that participate in research and increase the quality of science that students are exposed to in this region of the country.