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

Sporbert, Anje and Gahl, Anja and Ankerhold, Richard and Leonhardt, Heinrich and Cardoso, M Cristina (2002):
DNA polymerase clamp shows little turnover at established replication sites but sequential de novo assembly at adjacent origin clusters.
10, In: Molecular cell, (6), pp. 1355-65, ISSN 1097-2765, [Online-Edition: http://www.cardoso-lab.org/publications/Sporbert_2002+Suppl....],
[Article]

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.

Item Type: Article
Erschienen: 2002
Creators: Sporbert, Anje and Gahl, Anja and Ankerhold, Richard and Leonhardt, Heinrich and Cardoso, M Cristina
Title: DNA polymerase clamp shows little turnover at established replication sites but sequential de novo assembly at adjacent origin clusters.
Language: German
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.

Journal or Publication Title: Molecular cell
Volume: 10
Number: 6
Divisions: 10 Department of Biology > Cell Biology and Epigenetics
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10 Department of Biology
Date Deposited: 06 Mar 2010 07:21
Official URL: http://www.cardoso-lab.org/publications/Sporbert_2002+Suppl....
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