A Live-Cell Imaging Approach for Measuring DNA Replication Rates
Author(s) -
Daniel Dovrat,
Danielle Dahan,
Shachar Sherman,
Ioannis Tsirkas,
Natalie Elia,
Amir Aharoni
Publication year - 2018
Publication title -
cell reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.264
H-Index - 154
eISSN - 2639-1856
pISSN - 2211-1247
DOI - 10.1016/j.celrep.2018.06.018
Subject(s) - replication (statistics) , dna replication , live cell imaging , dna , microbiology and biotechnology , computational biology , biology , cell , genetics , virology
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.
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