title: Direct Evidence for the Formation of Precatenanes during DNA Replication.
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.
authors.affiliation: From the Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas, Ramiro de Maeztu 9, 28040, Madrid, Spain and.
authors.forename: Jorge
authors.initials: J
authors.lastname: Cebrián
authors.affiliation: From the Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas, Ramiro de Maeztu 9, 28040, Madrid, Spain and.
authors.forename: Alicia
authors.initials: A
authors.lastname: Castán
authors.affiliation: the Scientific and Applied Computing Laboratory, Polytechnic School, National University of Asunción. P.O. Box 2111 SL. San Lorenzo, Paraguay.
authors.forename: Víctor
authors.initials: V
authors.lastname: Martínez
authors.affiliation: the Scientific and Applied Computing Laboratory, Polytechnic School, National University of Asunción. P.O. Box 2111 SL. San Lorenzo, Paraguay.
authors.forename: Maridian J
authors.initials: MJ
authors.lastname: Kadomatsu-Hermosa
authors.affiliation: the Scientific and Applied Computing Laboratory, Polytechnic School, National University of Asunción. P.O. Box 2111 SL. San Lorenzo, Paraguay.
authors.forename: Cristina
authors.initials: C
authors.lastname: Parra
authors.affiliation: the Scientific and Applied Computing Laboratory, Polytechnic School, National University of Asunción. P.O. Box 2111 SL. San Lorenzo, Paraguay.
authors.forename: María José
authors.initials: MJ
authors.lastname: Fernández-Nestosa
authors.affiliation: the Scientific and Applied Computing Laboratory, Polytechnic School, National University of Asunción. P.O. Box 2111 SL. San Lorenzo, Paraguay.
authors.forename: Christian
authors.initials: C
authors.lastname: Schaerer
authors.affiliation: From the Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas, Ramiro de Maeztu 9, 28040, Madrid, Spain and.
authors.forename: Pablo
authors.initials: P
authors.lastname: Hernández
authors.affiliation: From the Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas, Ramiro de Maeztu 9, 28040, Madrid, Spain and.
authors.forename: Dora B
authors.initials: DB
authors.lastname: Krimer
authors.affiliation: From the Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas, Ramiro de Maeztu 9, 28040, Madrid, Spain and schvartzman@cib.csic.es.
authors.forename: Jorge B
authors.initials: JB
authors.lastname: Schvartzman
mesh_terms: D002384:Catalysis; D003470:Culture Media / Q000737:chemistry; D004261:DNA Replication; D027101:DNA Topoisomerase IV / Q000737:chemistry; D004269:DNA, Bacterial / Q000235:genetics*; D004278:DNA, Superhelical / Q000235:genetics; D015195:Drug Design; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000378:metabolism
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D004278:DNA, Superhelical; D027101:DNA Topoisomerase IV
keywords: DNA replication; DNA topoisomerase; DNA topology; catenation; cell biology; knotting; molecular biology; plasmid; supercoiling; topoisomerases
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