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Dynamics of DNA replication factories in living cells

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Item Type:Article
Title:Dynamics of DNA replication factories in living cells
Creators Name:Leonhardt, H. and Rahn, H.P. and Weinzierl, P. and Sporbert, A. and Cremer, T. and Zink, D. and Cardoso, M.C.
Abstract:DNA replication occurs in microscopically visible complexes at discrete sites (replication foci) in the nucleus. These foci consist of DNA associated with replication machineries, i.e., large protein complexes involved in DNA replication. To study the dynamics of these nuclear replication foci in living cells, we fused proliferating cell nuclear antigen (PCNA), a central component of the replication machinery, with the green fluorescent protein (GFP). Imaging of stable cell lines expressing low levels of GFP-PCNA showed that replication foci are heterogeneous in size and lifetime. Time-lapse studies revealed that replication foci clearly differ from nuclear speckles and coiled bodies as they neither show directional movements, nor do they seem to merge or divide. These four dimensional analyses suggested that replication factories are stably anchored in the nucleus and that changes in the pattern occur through gradual, coordinated, but asynchronous, assembly and disassembly throughout S phase.
Keywords:Cell Cycle, DNA Replication Foci, Green Fluorescent Protein, Nuclear Organization, Proliferating Cell Nuclear Antigen, Animals, Mice
Source:Journal of Cell Biology
ISSN:0021-9525
Publisher:Rockefeller University Press (U.S.A.)
Volume:149
Number:2
Page Range:271-279
Date:17 April 2000
Additional Information:Copyright (c) 2000 by The Rockefeller University Press
Official Publication:http://www.jcb.org/cgi/content/abstract/149/2/271
PubMed:View item in PubMed

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