Publication date: 3 July 2018
Source: Cell Reports, Volume 24, Issue 1
Author(s): Daniel Dovrat, Danielle Dahan, Shachar Sherman, Ioannis Tsirkas, Natalie Elia, Amir Aharoni
Summary
We describe a simple and direct approach to measure the progression of single DNA replication forks in living cells by monitoring two fluorescently labeled loci downstream of an origin of replication. We employ this approach to investigate the roles of several leading and lagging strand factors in overall replisome function and show that fork progression is strongly dependent on proper maturation of Okazaki fragments. We also demonstrate how related cellular phenotypes, such as cell-cycle progression and the dynamics of sister chromatid cohesion, may be simultaneously monitored and correlated to DNA replication at the single-cell level.
Graphical Abstract
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