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PMID: 10769021 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Dynamics of DNA replication factories in living cells.

The Journal of cell biology ·Vol. 149 ·No. 2 ·2000-04-17 ·Pages 271-80

Leonhardt H, Rahn HP, Weinzierl P, Sporbert A, Cremer T, Zink D, Cardoso MC

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.

MeSH Terms
Animals COS Cells Cell Cycle/physiology Cell Line Cell Nucleus/physiology,ultrastructure DNA Replication Green Fluorescent Proteins Humans Kinetics Luminescent Proteins/analysis,genetics Mice Microscopy, Video/methods Muscle, Skeletal Proliferating Cell Nuclear Antigen/genetics,metabolism Recombinant Fusion Proteins/metabolism Transfection
Chemicals
Luminescent Proteins Proliferating Cell Nuclear Antigen Recombinant Fusion Proteins Green Fluorescent Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Leonhardt H
Max Delbrück Center for Molecular Medicine, D-13125 Berlin, Germany.
Rahn H P
Weinzierl P
Sporbert A
Cremer T
Zink D
Cardoso M C
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.149.2.271
Published
2000-04-17
Pages
271-80
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2175147
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.149.2.271 PMC Full Text View journal details
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