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First published online 5 October 2004
doi: 10.1242/jcs.01412

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Stable chromosomal units determine the spatial and temporal organization of DNA replication

¹ University of Munich (LMU), Department Biology II, Grosshaderner Str. 2, 82152 Planegg-Martinsried, Germany
² Max Delbrück Center of Molecular Medicine, Wiltbergstr. 50, 13125 Berlin, Germany
³ European Molecular Biology Laboratory (EMBL), Meyerhofstr. 1, 69117 Heidelberg, Germany

^* Author for correspondence (e-mail: dani.zink{at}lrz.uni-muenchen.de)

Accepted 19 July 2004

DNA replication occurs in mammalian cells at so-called replication foci occupying defined nuclear sites at specific times during S phase. It is an unresolved problem how this specific spatiotemporal organization of replication foci is determined. Another unresolved question remains as to what extent DNA is redistributed during S phase. To investigate these problems, we visualized the replicating DNA and the replication machinery simultaneously in living HeLa cells. Time-lapse analyses revealed that DNA was not redistributed to other nuclear sites during S phase. Furthermore, the results showed that DNA is organized into stable aggregates equivalent to replication foci. These aggregates, which we call sub-chromosomal foci, stably maintained their replication timing from S phase to S phase. During S-phase progression, the replication machinery sequentially proceeded through spatially adjacent sets of sub-chromosomal foci. These findings imply that the specific nuclear substructure of chromosomes and the order of their stable subunits determine the spatiotemporal organization of DNA replication.

Key words: Replication dynamics, Replication foci, Sub-chromosomal foci

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