Aviso: para depositar documentos, por favor, inicia sesión e identifícate con tu cuenta de correo institucional de la UCM con el botón MI CUENTA UCM. No emplees la opción AUTENTICACIÓN CON CONTRASEÑA Disculpen las molestias.
 

Two-Dimensional Gel Electrophoresis to Study the Activity of Type IIA Topoisomerases on Plasmid Replication Intermediates

dc.contributor.authorCebrián Castillo, Jorge
dc.contributor.authorMartínez, Victor
dc.contributor.authorHernández, Pablo
dc.contributor.authorKrimer, Dora B.
dc.contributor.authorFernández Nestosa, María José
dc.contributor.authorSchvartzman, Jorge B.
dc.date.accessioned2023-06-16T14:20:28Z
dc.date.available2023-06-16T14:20:28Z
dc.date.issued2021-11-13
dc.description.abstractDNA topoisomerases are the enzymes that regulate DNA topology in all living cells. Since the discovery and purification of ω (omega), when the first were topoisomerase identified, the function of many topoisomerases has been examined. However, their ability to relax supercoiling and unlink the pre-catenanes of partially replicated molecules has received little attention. Here, we used two-dimensional agarose gel electrophoresis to test the function of three type II DNA topoisomerases in vitro: the prokaryotic DNA gyrase, topoisomerase IV and the human topoisomerase 2α. We examined the proficiency of these topoisomerases on a partially replicated bacterial plasmid: pBR-TerE@AatII, with an unidirectional replicating fork, stalled when approximately half of the plasmid had been replicated in vivo. DNA was isolated from two strains of Escherichia coli: DH5αF’ and parE10. These experiments allowed us to assess, for the first time, the efficiency of the topoisomerases examined to resolve supercoiling and pre-catenanes in partially replicated molecules and fully replicated catenanes formed in vivo. The results obtained revealed the preferential functions and also some redundancy in the abilities of these DNA topoisomerases in vitro.
dc.description.departmentDepto. de Farmacología y Toxicología
dc.description.facultyFac. de Medicina
dc.description.refereedTRUE
dc.description.sponsorshipMinisterio de Economía, Industria y Competitividad (MINECO)
dc.description.sponsorshipCONACYT-PROCIENCIA
dc.description.statuspub
dc.eprint.idhttps://eprints.ucm.es/id/eprint/70761
dc.identifier.doi10.3390/biology10111195
dc.identifier.issn2079-7737
dc.identifier.officialurlhttps://doi.org/10.3390/biology10111195
dc.identifier.relatedurlhttps://www.mdpi.com/2079-7737/10/11/1195/htm
dc.identifier.urihttps://hdl.handle.net/20.500.14352/4744
dc.issue.number11
dc.journal.titleBiology
dc.language.isoeng
dc.page.initial1195
dc.publisherMDPI
dc.relation.projectIDBFU2014-56835
dc.relation.projectIDPINV15-573
dc.rightsAtribución 3.0 España
dc.rights.accessRightsopen access
dc.rights.urihttps://creativecommons.org/licenses/by/3.0/es/
dc.subject.keywordDNA topology
dc.subject.keywordreplication
dc.subject.keywordsupercoiling
dc.subject.keywordpre-catenation
dc.subject.keywordtwo-dimensional agarose gel electrophoresis
dc.subject.ucmGenética médica
dc.subject.unesco2410.07 Genética Humana
dc.titleTwo-Dimensional Gel Electrophoresis to Study the Activity of Type IIA Topoisomerases on Plasmid Replication Intermediates
dc.typejournal article
dc.volume.number10
dspace.entity.typePublication
relation.isAuthorOfPublication99808a19-fe45-4a53-a877-d83b03153d0b
relation.isAuthorOfPublication.latestForDiscovery99808a19-fe45-4a53-a877-d83b03153d0b

Download

Original bundle

Now showing 1 - 1 of 1
Loading...
Thumbnail Image
Name:
biology-10-01195.pdf
Size:
1.75 MB
Format:
Adobe Portable Document Format

Collections