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4D Visualization of replication foci in mammalian cells corresponding to individual replicons

Abstract:
Since the pioneering proposal of the replicon model of DNA replication 50 years ago, the predicted replicons have not been identified and quantified at the cellular level. Here, we combine conventional and super-resolution microscopy of replication sites in live and fixed cells with computational image analysis. We complement these data with genome size measurements, comprehensive analysis of S-phase dynamics and quantification of replication fork speed and replicon size in human and mouse cells. These multidimensional analyses demonstrate that replication foci (RFi) in three-dimensional (3D) preserved somatic mammalian cells can be optically resolved down to single replicons throughout S-phase. This challenges the conventional interpretation of nuclear RFi as replication factories, that is, the complex entities that process multiple clustered replicons. Accordingly, 3D genome organization and duplication can be now followed within the chromatin context at the level of individual replicons.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1038/ncomms11231

Authors


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Institution:
University of Oxford
Division:
MSD
Department:
Biochemistry
Role:
Author


More from this funder
Funding agency for:
Cardoso, M
Grant:
DFG CA 198/9
More from this funder
Funding agency for:
Leonhardt, H
Grant:
DFG SFB 1064 A17/Z05
More from this funder
Funding agency for:
Schermelleh, L
Leonhardt, H
Cardoso, M
Grant:
DFG SFB-TR5
DFG SFB 1064 A17/Z05
DFG CA 198/9


Publisher:
Nature Publishing Group
Journal:
Nature Communications More from this journal
Volume:
7
Pages:
11231
Publication date:
2016-04-07
Acceptance date:
2016-03-03
DOI:
EISSN:
2041-1723


Pubs id:
pubs:614778
UUID:
uuid:52fa8365-00c6-4761-9b40-3ad3b608b4b1
Local pid:
pubs:614778
Source identifiers:
614778
Deposit date:
2016-04-10

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