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

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Official URL:https://doi.org/10.1038/ncomms11231
PubMed:View item in PubMed
Creators Name:Chagin, V.O. and Casas-Delucchi, C.S. and Reinhart, M. and Schermelleh, L. and Markaki, Y. and Maiser, A. and Bolius, J.J. and Bensimon, A. and Fillies, M. and Domaing, P. and Rozanov, Y.M. and Leonhardt, H. and Cardoso, M.C.
Journal Title:Nature Communications
Journal Abbreviation:Nat Commun
Volume:7
Page Range:11231
Date:7 April 2016
Keywords:Cell Line, Chromatin, Computer-Assisted Image Processing, DNA Replication, Gene Expression, Genome Size, HeLa Cells, Kinetics, Molecular Imaging, Myoblasts, Proliferating Cell Nuclear Antigen, Replicon, S Phase, Species Specificity, Animals, Mice
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.
ISSN:2041-1723
Publisher:Nature Publishing Group (U.K.)
Item Type:Article

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