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

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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.