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• W2022131374 abstract "The dynamics of DNA topology during replication are still poorly understood. Bacterial plasmids are negatively supercoiled. This underwinding facilitates strand separation of the DNA duplex during replication. Leading the replisome, a DNA helicase separates the parental strands that are to be used as templates. This strand separation causes overwinding of the duplex ahead. If this overwinding persists, it would eventually impede fork progression. In bacteria, DNA gyrase and topoisomerase IV act ahead of the fork to keep DNA underwound. However, the processivity of the DNA helicase might overcome DNA gyrase and topoisomerase IV. It was proposed that the overwinding that builds up ahead of the fork could force it to swivel and diffuse this positive supercoiling behind the fork where topoisomerase IV would also act to maintain replicating the DNA underwound. Putative intertwining of sister duplexes in the replicated region are called precatenanes. Fork swiveling and the formation of precatenanes, however, are still questioned. Here, we used classical genetics and high resolution two-dimensional agarose gel electrophoresis to examine the torsional tension of replication intermediates of three bacterial plasmids with the fork stalled at different sites before termination. The results obtained indicated that precatenanes do form as replication progresses before termination.Background: Changes in DNA topology during replication are still poorly understood.Results: Classical genetics and two-dimensional agarose gel electrophoresis showed that RIs tensioned in the absence of Topo IV.Conclusion: The results indicated that replication forks swivel in vivo leading to the formation of precatenanes as replication progresses.Significance: This conclusion ends a long lasting debate on the formation of precatenanes during replication. The dynamics of DNA topology during replication are still poorly understood. Bacterial plasmids are negatively supercoiled. This underwinding facilitates strand separation of the DNA duplex during replication. Leading the replisome, a DNA helicase separates the parental strands that are to be used as templates. This strand separation causes overwinding of the duplex ahead. If this overwinding persists, it would eventually impede fork progression. In bacteria, DNA gyrase and topoisomerase IV act ahead of the fork to keep DNA underwound. However, the processivity of the DNA helicase might overcome DNA gyrase and topoisomerase IV. It was proposed that the overwinding that builds up ahead of the fork could force it to swivel and diffuse this positive supercoiling behind the fork where topoisomerase IV would also act to maintain replicating the DNA underwound. Putative intertwining of sister duplexes in the replicated region are called precatenanes. Fork swiveling and the formation of precatenanes, however, are still questioned. Here, we used classical genetics and high resolution two-dimensional agarose gel electrophoresis to examine the torsional tension of replication intermediates of three bacterial plasmids with the fork stalled at different sites before termination. The results obtained indicated that precatenanes do form as replication progresses before termination. Background: Changes in DNA topology during replication are still poorly understood. Results: Classical genetics and two-dimensional agarose gel electrophoresis showed that RIs tensioned in the absence of Topo IV. Conclusion: The results indicated that replication forks swivel in vivo leading to the formation of precatenanes as replication progresses. Significance: This conclusion ends a long lasting debate on the formation of precatenanes during replication." @default.
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• W2022131374 date "2015-05-01" @default.
• W2022131374 modified "2023-09-26" @default.
• W2022131374 title "Direct Evidence for the Formation of Precatenanes during DNA Replication" @default.
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• W2022131374 doi "https://doi.org/10.1074/jbc.m115.642272" @default.
• W2022131374 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4447951" @default.
• W2022131374 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/25829493" @default.
• W2022131374 hasPublicationYear "2015" @default.
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