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DNA polymerase clamp shows little turnover at established replication sites but sequential de novo assembly at adjacent origin clusters

Official URL:https://doi.org/10.1016/S1097-2765(02)00729-3
PubMed:View item in PubMed
Creators Name:Sporbert, A. and Gahl, A. and Ankerhold, R. and Leonhardt, H. and Cardoso, M.C.
Journal Title:Molecular Cell
Journal Abbreviation:Mol Cell
Page Range:1355-1365
Date:December 2002
Keywords:Cell Line, Chromatin, DNA Replication, DNA-Directed DNA Polymerase, Green Fluorescent Proteins, Kinetics, Luminescent Proteins, Myoblasts, Proliferating Cell Nuclear Antigen, Recombinant Fusion Proteins, Replication Origin, Signal Transduction, Animals, Mice
Abstract:The spatial and temporal organization of DNA replication was investigated in living cells with a green fluorescent protein fusion to the DNA polymerase clamp PCNA. In situ extractions and photobleaching experiments revealed that PCNA, unlike RPA34, shows little if any turnover at replication sites, suggesting that it remains associated with the replication machinery through multiple rounds of Okazaki fragment synthesis. Photobleaching analyses further showed that the transition from earlier to later replicons occurs by disassembly into a nucleoplasmic pool of rapidly diffusing subcomponents and reassembly at newly activated sites. The fact that these replication sites were de novo assembled in close proximity to earlier ones suggests that activation of neighboring origins may occur by a domino effect possibly involving local changes in chromatin structure and accessibility.
Publisher:Cell Press (U.S.A.)
Item Type:Article

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