A Live-Cell Imaging Approach for Measuring DNA Replication Rates

Daniel Dovrat; Danielle Dahan; Shachar Sherman; Ioannis Tsirkas; Natalie Elia; Amir Aharoni

doi:10.1016/j.celrep.2018.06.018

A Live-Cell Imaging Approach for Measuring DNA Replication Rates

Daniel Dovrat, Danielle Dahan, Shachar Sherman, Ioannis Tsirkas, Natalie Elia, Amir Aharoni

Department of Life Sciences

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	252-258
Number of pages	7
Journal	Cell Reports
Volume	24
Issue number	1
DOIs	https://doi.org/10.1016/j.celrep.2018.06.018
State	Published - 3 Jul 2018

Keywords

DNA replication
FROS
Okazaki fragment maturation
live-cell imaging
sister chromatid cohesion

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/j.celrep.2018.06.018

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title = "A Live-Cell Imaging Approach for Measuring DNA Replication Rates",

abstract = "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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AU - Dovrat, Daniel

AU - Dahan, Danielle

AU - Sherman, Shachar

AU - Tsirkas, Ioannis

AU - Elia, Natalie

AU - Aharoni, Amir

PY - 2018/7/3

Y1 - 2018/7/3

N2 - 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.

AB - 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.

KW - DNA replication

KW - FROS

KW - Okazaki fragment maturation

KW - live-cell imaging

KW - sister chromatid cohesion

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SP - 252

EP - 258

JO - Cell Reports

JF - Cell Reports

IS - 1

ER -

A Live-Cell Imaging Approach for Measuring DNA Replication Rates

Abstract

Keywords

ASJC Scopus subject areas

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