CAREER: Under the Hood of the Replisome: Single-molecule Mechanics of DNA Replication

Harvard University, Cambridge MA

Investigators

Abstract

The research component of this project focuses on the development and utilization of single molecule techniques to study the complex processes involved in DNA replication. The bacteriophage T7 DNA replication machinery will be examined. The mechanical manipulation of individual DNA molecules will be combined with the imaging of single fluorescently labeled proteins to study how double-stranded DNA is unwound and how this unwinding is coupled to DNA synthesis. By making 'molecular movies' of these processes at the single-molecule level, details and events can be observed that would otherwise be hidden under the ensemble average. The education component of this proposal strives to bridge the gap that exists between the physics education offered to biology graduate students and the increasing importance of quantitative thinking in biological research. A novel course will be developed that aims to train biology graduate and upper-level undergraduate students in the physical principles that are important in biological processes at the molecular scale. Interactive web-based, 'virtual' experiments will be created to let students develop an intuitive feeling for the role of physics in the behavior of biological macromolecules. The lecture materials will be converted into a textbook that is accompanied by a CD-ROM containing the interactive, virtual experiments.