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

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. Cristina (2016):
4D Visualization of replication foci in mammalian cells corresponding to individual replicons.
In: Nature communications, p. 11231, 7, ISSN 2041-1723,
[Article]

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.

Item Type: Article
Erschienen: 2016
Creators: 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. Cristina
Title: 4D Visualization of replication foci in mammalian cells corresponding to individual replicons.
Language: English
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.

Journal or Publication Title: Nature communications
Volume: 7
Divisions: 10 Department of Biology
10 Department of Biology > Cell Biology and Epigenetics
Date Deposited: 19 Apr 2016 10:16
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