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

V. O. Chagin, C. S. Casas-Delucchi, M. Reinhart, L. Schermelleh, Y. Markaki, A. Maiser, J. J. Bolius, A. Bensimon, M. Fillies, P. Domaing, Y. M. Rozanov, H. Leonhardt and M. C. Cardoso ()
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V. O. Chagin: Technische Universitaet Darmstadt
C. S. Casas-Delucchi: Technische Universitaet Darmstadt
M. Reinhart: Technische Universitaet Darmstadt
L. Schermelleh: Micron Advanced Bioimaging Unit, University of Oxford
Y. Markaki: Ludwig Maximilians University Munich
A. Maiser: Ludwig Maximilians University Munich
J. J. Bolius: Technische Universitaet Darmstadt
A. Bensimon: Genomic Vision
M. Fillies: Max Delbrueck Center for Molecular Medicine
P. Domaing: Max Delbrueck Center for Molecular Medicine
Y. M. Rozanov: Laboratory of chromosome stability, Institute of Cytology
H. Leonhardt: Ludwig Maximilians University Munich
M. C. Cardoso: Technische Universitaet Darmstadt

Nature Communications, 2016, vol. 7, issue 1, 1-12

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms11231

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