Year

2019

Degree Name

Doctor of Philosophy

Department

School of Chemistry

Abstract

The replisome is a complex, multi-protein machinery that copies genomic DNA before cell division and allows the faithful transmission of genetic information to the next generation. Error-free duplication is achieved through a fine-tuned coordination among the various components of the replisome. Biochemical and biophysical techniques have contributed tremendously to exposing the role of each component and interactions within the replisome. The recent introduction of single-molecule approaches has revolutionized our understanding of complex protein systems by allowing access to molecular dynamics without the need for population averaging. Advances in improving the throughput of single-molecule techniques allow researchers to reliably sample subpopulations and follow rare events to elucidate behaviour of multi-protein systems that is much more heterogeneous and stochastic than thought before. This thesis reveals the molecular dynamics at the replication fork in Escherichia coli, exploiting the kinetic resolution of single-molecule imaging techniques combined with the diversity of biochemical ensemble-averaging approaches.

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Unless otherwise indicated, the views expressed in this thesis are those of the author and do not necessarily represent the views of the University of Wollongong.