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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.

Original publication

DOI

10.1038/ncomms11231

Type

Journal article

Journal

Nat Commun

Publication Date

07/04/2016

Volume

7

Keywords

Animals, Cell Line, Chromatin, DNA Replication, Gene Expression, Genome Size, HeLa Cells, Humans, Image Processing, Computer-Assisted, Kinetics, Mice, Molecular Imaging, Myoblasts, Proliferating Cell Nuclear Antigen, Replicon, S Phase, Species Specificity