CAREER: High-throughput Single-molecule Studies of Replication Conflict and Restart.

University Of Texas At Austin, Austin TX

Investigators

Abstract

The genomes of all living organisms must complete two essential functions-DNA replication and transcription. Conflicts between the replication and transcription machineries are pervasive on DNA and can lead to catastrophic outcomes including structural damage to DNA and mutagenesis. The goal of this project is to examine at a molecular level how replication-transcription conflicts occur and are resolved. The exciting research topic of "molecular machines" will also be used to inspire students' interest in science at the early-education stage. As part of the institutional Freshman Research Initiative undergraduates with minimal laboratory experience will be trained in inquiry-based research using quantitative biochemical and biophysical methods. Graduate students and postdoctoral fellows will be trained in these interdisciplinary areas as well. Additionally, the PI will participate in projects aimed at exposing local under-resourced middle school students to research early in their education. High-throughput single-molecule fluorescence imaging will be used to directly observe the intricate choreography of replication and transcription enzymes as they encounter each other on the same DNA molecule. The findings will shed light on replication-transcription conflicts, and help resolve the larger problem of how organisms replicate a genome that is crowded with many DNA-binding proteins. As part of the project, new methods for high-throughput single-molecule biology will be developed by combining elements of micro-fabrication and surface chemistry, and disseminated to the research community. The project is co-funded by the Directorate for Biological Sciences (Division of Molecular and Cellular Biosciences: Genetic Mechanisms program and the Division of Emerging Frontiers) and by the Directorate for Mathematical and Physical Sciences (Division of Chemistry: Chemistry of Life Sciences program).