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Visualization of replication conflicts and other stories of life at a molecular scale - Applications of the 2014 Nobel Prize in Chemistry

Paul Wiggins, University of Washington, Departments of Physics and Bioengineering
Monday, October 27, 2014 - 4:00pm to 5:00pm
PAA A-102
The 2014 Nobel Prize in Chemistry was awarded for the development of Super-Resolution Fluorescence Microscopy. The resolution of traditional wide-field fluorescence microscopy is limited by diffraction to roughly 200 nm, yet the scale of the molecular building blocks of life are two orders of magnitude smaller. Super-resolution techniques currently increase the resolution of fluorescence microscopy by a factor of ten in many practical in vivo biological applications. This increase in resolution has the potential to provide a great many new insights into a very wide-range of biological phenomena in which cellular function and ultrastructure are intimately linked.

In this talk, I will briefly discuss the physics behind the prize, before presenting new results from my lab on DNA replication (and other stories) using techniques directly inspired by the work of the Nobel Laureates. Cell proliferation requires timely and reliable DNA replication. Genetic and biochemical evidence reveals that the replication process is subject to a variety of conflict mechanisms, including DNA damage, concurrent DNA transcription, and DNA-bound protein complexes which can all act to stall the replication process. Without a mechanism for rapid and efficient resolution, these conflicts have the potential to cause genomic instability and even cell death. The frequency of these conflicts and their consequences to replisome structure remain unknown. We report the direct visualization of single replication conflicts with single-molecule sensitivity in two model organisms well suited for single-molecule microscopy (E. coli and Bacillus subtilis) and characterize the dynamics of these conflicts in vivo.

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