← Back to Search
Live-Cell Imaging Reveals Replication Of Individual Replicons In Eukaryotic Replication Factories
E. Kitamura, J. Blow, T. Tanaka
Published 2006 · Medicine, Biology
Download PDFAnalyze on Scholarcy
Faithful DNA replication ensures genetic integrity in eukaryotic cells, but it is still obscure how replication is organized in space and time within the nucleus. Using timelapse microscopy, we have developed a new assay to analyze the dynamics of DNA replication both spatially and temporally in individual Saccharomyces cerevisiae cells. This allowed us to visualize replication factories, nuclear foci consisting of replication proteins where the bulk of DNA synthesis occurs. We show that the formation of replication factories is a consequence of DNA replication itself. Our analyses of replication at specific DNA sequences support a long-standing hypothesis that sister replication forks generated from the same origin stay associated with each other within a replication factory while the entire replicon is replicated. This assay system allows replication to be studied at extremely high temporal resolution in individual cells, thereby opening a window into how replication dynamics vary from cell to cell.
This paper references
Is the yeast Anaphase Promoting Complex needed to prevent re‐replication during G2 and M phases?
S. Pichler (1997)
Early events in DNA replication require cyclin E and are blocked by p21CIP1
P. Jackson (1995)
Methods in Yeast Genetics
M. Rose (1990)
Flexibility and governance in eukaryotic DNA replication.
E. Schwob (2004)
Xenopus cdc7 function is dependent on licensing but not on XORC, XCdc6, or CDK activity and is required for XCdc45 loading.
P. Jares (2000)
Putting the HO gene to work: practical uses for mating-type switching.
I. Herskowitz (1991)
Eukaryotic DNA Replication
T. Kelly (1988)
Termination of DNA replication of bacterial and plasmid chromosomes
D. Bussiere (1999)
Molecular mechanisms of kinetochore capture by spindle microtubules
K. Tanaka (2005)
DNA polymerases α, δ, and ɛ localize and function together at replication forks in Saccharomyces cerevisiae
Shin-ichiro Hiraga (2005)
GFP tagging of budding yeast chromosomes reveals that protein–protein interactions can mediate sister chromatid cohesion
A. Straight (1996)
S-phase-promoting cyclin-dependent kinases prevent re-replication by inhibiting the transition of replication origins to a pre-replicative state
Christian Dahmann (1995)
Cellular DNA replicases: components and dynamics at the replication fork.
A. Johnson (2005)
Targeting of PCNA to sites of DNA replication in the mammalian cell nucleus *
S. Somanathan (2001)
Cohesion between sister chromatids must be established during DNA replication
F. Uhlmann (1998)
Activation of S-phase-promoting CDKs in late G1 defines a "point of no return" after which Cdc6 synthesis cannot promote DNA replication in yeast.
S. Piatti (1996)
Monitoring S phase progression globally and locally using BrdU incorporation in TK(+) yeast strains.
A. Lengronne (2001)
DNA polymerase clamp shows little turnover at established replication sites but sequential de novo assembly at adjacent origin clusters.
A. Sporbert (2002)
DNA replication: a complex matter
I. Frouin (2003)
Reconstitution of an efficient thymidine salvage pathway in Saccharomyces cerevisiae.
Laurence Vernis (2003)
The chromosome cycle: coordinating replication and segregation
J. Blow (2005)
Central role for Cdc45 in establishing an initiation complex of DNA replication in Xenopus egg extracts
S. Mimura (2000)
Mcm2, but Not Rpa, Is a Component of the Mammalian Early G1-Phase Prereplication Complex
D. S. Dimitrova (1999)
Right Place, Right Time, and Only Once: Replication Initiation in Metazoans
Y. Machida (2005)
Association of RPA with chromosomal replication origins requires an Mcm protein, and is regulated by Rad53, and cyclin‐ and Dbf4‐dependent kinases
T. Tanaka (1998)
Cdc6 is an unstable protein whose de novo synthesis in G1 is important for the onset of S phase and for preventing a ‘reductional’ anaphase in the budding yeast Saccharomyces cerevisiae.
S. Piatti (1995)
The midcell replication factory in Bacillus subtilis is highly mobile: implications for coordinating chromosome replication with other cell cycle events
Margaret D Migocki (2004)
Spatial and temporal organization of replicating Escherichia coli chromosomes
I. Lau (2003)
The DNA Polymerase Domain of polε Is Required for Rapid, Efficient, and Highly Accurate Chromosomal DNA Replication, Telomere Length Maintenance, and Normal Cell Senescence inSaccharomyces cerevisiae *
Tomoko Ohya (2002)
Preventing re-replication of chromosomal DNA
J. Blow (2005)
Organization of DNA into foci during replication.
J. Newport (1996)
CLB5 and CLB6, a new pair of B cyclins involved in DNA replication in Saccharomyces cerevisiae.
E. Schwob (1993)
Polymerases and the Replisome: Machines within Machines
T. A. Baker (1998)
The DNA replication fork in eukaryotic cells.
S. Waga (1998)
Yeast DNA polymerases and their role at the replication fork.
A. Sugino (1995)
Point of No Return
J. Marquand (1949)
Chromosome Dynamics in the Yeast Interphase Nucleus
Structural organizations of replicon domains during DNA synthetic phase in the mammalian nucleus.
H. Nakamura (1986)
Simian virus 40 T-antigen DNA helicase is a hexamer which forms a binary complex during bidirectional unwinding from the viral origin of DNA replication.
R. Wessel (1992)
Dynamics of DNA Replication Factories in Living Cells
H. Leonhardt (2000)
Loading of an Mcm Protein onto DNA Replication Origins Is Regulated by Cdc6p and CDKs
T. Tanaka (1997)
A moving DNA replication factory in Caulobacter crescentus
R. B. Jensen (2001)
Eukaryotic DNA replication: a model for a fixed double replisome.
A. Falaschi (2000)
ORC-dependent and origin-specific initiation of DNA replication at defined foci in isolated yeast nuclei.
P. Pasero (1997)
A Mec1- and Rad53-dependent checkpoint controls late-firing origins of DNA replication
C. Santocanale (1998)
Splitting the chromosome: cutting the ties that bind sister chromatids.
K. Nasmyth (2001)
Visualization of replication factories attached to a nucleoskeleton
P. Hozák (1993)
Bidirectional chromosome replication: some topological considerations.
C. Dingman (1974)
A novel strategy for constructing N‐terminal chromosomal fusions to green fluorescent protein in the yeast Saccharomyces cerevisiae
B. Prein (2000)
Heterogeneity of eukaryotic replicons, replicon clusters, and replication foci
R. Berezney (2000)
Mating type–dependent constraints on the mobility of the left arm of yeast chromosome III
D. Bressan (2004)
Regulation of Chromatin Structure and Function
A. Wolffe (1994)
Temporal separation of replication and recombination requires the intra-S checkpoint
P. Meister (2005)
Movement of replicating DNA through a stationary replisome.
K. Lemon (2000)
Yeast Cdk1 translocates to the plus end of cytoplasmic microtubules to regulate bud cortex interactions
H. Maekawa (2003)
Localization of bacterial DNA polymerase: evidence for a factory model of replication.
K. Lemon (1998)
Chromatin Structure and Function
C. Nicolini (1979)
Replication dynamics of the yeast genome.
M. Raghuraman (2001)
Methods in Yeast Genetics (New York: CSHL press)
D. C. Amberg (2005)
SV40 DNA replication.
T. J. Kelly (1988)
AC Tose (1993)
Cohesins: Chromosomal Proteins that Prevent Premature Separation of Sister Chromatids
C. Michaelis (1997)
This paper is referenced by
Transcription and replication
A. Poveda (2010)
Targeted INO80 enhances subnuclear chromatin movement and ectopic homologous recombination.
Frank R Neumann (2012)
Investigation of the essential role of S. cerevisiae MEC1
Caroline Earp (2008)
Epigenetic regulation of higher order chromatin conformations and networks
Chengxi Shi (2013)
Replication fork stalling by bulky DNA damage: localization at active origins and checkpoint modulation
Eugen C. Minca (2011)
How and why multiple MCMs are loaded at origins of DNA replication
S. Das (2016)
The yeast genome undergoes significant topological reorganization in quiescence
Mark T Rutledge (2015)
Global regulation of genome duplication in eukaryotes: an overview from the epifluorescence microscope
J. Herrick (2007)
Comparative 3D Genome Structure Analysis of the Fission and the Budding Yeast
K. Gong (2015)
New insights into replisome fluidity during chromosome replication.
I. Kurth (2013)
Independent Positioning and Action of Escherichia coli Replisomes in Live Cells
R. Reyes-Lamothe (2008)
Viral DNA Replication-Dependent DNA Damage Response Activation during BK Polyomavirus Infection
Brandy Verhalen (2015)
Genetic instability is prevented by Mrc1-dependent spatio-temporal separation of replicative and repair activities of homologous recombination
F. Prado (2014)
Actin' between phase separated domains for heterochromatin repair.
Chetan C. Rawal (2019)
Organization of sister origins and replisomes during multifork DNA replication in Escherichia coli
S. Fossum (2007)
Kaposi’s Sarcoma-Associated Herpesvirus Genome Replication, Partitioning, and Maintenance in Latency
E. Ohsaki (2012)
The S phase checkpoint promotes the Smc5/6 complex dependent SUMOylation of Pol2, the catalytic subunit of DNA polymerase ε
Alicja Winczura (2019)
Chromosome Duplication in Saccharomyces cerevisiae
S. Bell (2016)
Chromatin Architectural Factors as Safeguards against Excessive Supercoiling during DNA Replication
S. M. Ahmed (2020)
the SM-phase transition A topoisomerase II-dependent mechanism for resetting replicons at
O. Cuvier (2008)
Replication fork movement sets chromatin loop size and origin choice in mammalian cells
Sylvain Courbet (2008)
Actively Replicating Domains Randomly Associate into Replication Factories
Jens Karschau (2015)
Blossom of CRISPR technologies and applications in disease treatment
Huayi Liu (2018)
Unravelling global genome organization by 3C-seq.
Hideki Tanizawa (2012)
DNA in motion during double-strand break repair.
Judith Miné-Hattab (2013)
The budding yeast nucleus.
A. Taddei (2010)
Replication timing and its emergence from stochastic processes.
J. Bechhoefer (2012)
Compartmentalization of the cell nucleus and spatial organization of the genome
A. Gavrilov (2015)
DNA replication: failures and inverted fusions.
A. Carr (2011)
A Three-Dimensional Model of the Yeast Genome
Z. Duan (2010)
Cell cycle-dependent kinetochore localization of condensin complex in Saccharomyces cerevisiae.
Sophie Bachellier-Bassi (2008)
Localization of recombination proteins and Srs2 reveals anti-recombinase function in vivo
R. Burgess (2009)See more