Single-molecule studies of Saccharomyces cerevisiae DNA replication

DNA replication in eukaryotic organisms is a highly intricate process involving a large number of functionally and structurally distinct proteins. Despite being significantly different from DNA replication in bacterial and viral systems, eukaryotic DNA replication mechanisms have been found to be highly conserved across all eukaryotes, from yeast to humans. The budding yeast, Saccharomyces cerevisiae, has emerged as an important model organism to understand DNA replication in eukaryotes. This thesis describes the development and application of single-molecule approaches to study DNA replication in Saccharomyces cerevisiae. DNA replication reactions were reconstituted using purified protein factors and model DNA substrates on imaging platforms that enable us to observe and probe these mechanisms at the single-molecule level. Single-molecule techniques are designed for the observation of distinct DNA replication events and distinct replication factors, leading to an in-depth understanding of biomolecular mechanisms, without shielding variable behaviours within a population.