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dc.contributor.authorSpinks, Richard R
dc.contributor.authorSpenkelink, Lisanne M
dc.contributor.authorStratmann, Sarah A
dc.contributor.authorXu, Zhi-Qiang
dc.contributor.authorStamford, N Patrick J
dc.contributor.authorBrown, Susan E
dc.contributor.authorDixon, Nicholas E.
dc.contributor.authorJergic, Slobodan
dc.contributor.authorvan Oijen, Antoine M
dc.date.accessioned2021-09-08T06:51:43Z
dc.date.available2021-09-08T06:51:43Z
dc.date.issued2021-06-17
dc.identifier.citationSpinks, R. R., Spenkelink, L. M., Stratmann, S. A., Xu, Z.-Q., Stamford, N. P. J., Brown, S. E., … van Oijen, A. M. (2021). DnaB helicase dynamics in bacterial DNA replication resolved by single-molecule studies. Nucleic Acids Research, 49(12), 6804–6816. doi:10.1093/nar/gkab493
dc.identifier.issn0305-1048
dc.identifier.issn0305-1048
dc.identifier.pmid34139009
dc.identifier.doi10.1093/nar/gkab493
dc.identifier.urihttp://hdl.handle.net/10754/671115
dc.description.abstractIn Escherichia coli, the DnaB helicase forms the basis for the assembly of the DNA replication complex. The stability of DnaB at the replication fork is likely important for successful replication initiation and progression. Single-molecule experiments have significantly changed the classical model of highly stable replication machines by showing that components exchange with free molecules from the environment. However, due to technical limitations, accurate assessments of DnaB stability in the context of replication are lacking. Using in vitro fluorescence single-molecule imaging, we visualise DnaB loaded on forked DNA templates. That these helicases are highly stable at replication forks, indicated by their observed dwell time of ∼30 min. Addition of the remaining replication factors results in a single DnaB helicase integrated as part of an active replisome. In contrast to the dynamic behaviour of other replisome components, DnaB is maintained within the replisome for the entirety of the replication process. Interestingly, we observe a transient interaction of additional helicases with the replication fork. This interaction is dependent on the τ subunit of the clamp-loader complex. Collectively, our single-molecule observations solidify the role of the DnaB helicase as the stable anchor of the replisome, but also reveal its capacity for dynamic interactions.
dc.description.sponsorshipAustralian Research Council [DP150100956, DP180100858 to A.M.v.O., N.E.D.]; Australian Laureate Fellowship [FL140100027 to A.M.v.O.]; King Abdullah University of Science and Technology, Saudi Arabia [OSR-2015-CRG4-2644 to N.E.D., A.M.v.O.]; Australian Government Research Training Program Scholarship (to R.R.S). Funding for open access charge: Australian Research Council.
dc.publisherOxford University Press (OUP)
dc.relation.urlhttps://academic.oup.com/nar/article/49/12/6804/6303457
dc.rightsThis is a pre-copyedited, author-produced PDF of an article accepted for publication in Nucleic acids research following peer review. The version of record is available online at: https://academic.oup.com/nar/article/49/12/6804/6303457.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.titleDnaB helicase dynamics in bacterial DNA replication resolved by single-molecule studies.
dc.typeArticle
dc.identifier.journalNucleic acids research
dc.identifier.pmcidPMC8266626
dc.eprint.versionPublisher's Version/PDF
dc.contributor.institutionMolecular Horizons and School of Chemistry and Molecular Bioscience, University of Wollongong, Wollongong, New South Wales 2522, Australia
dc.contributor.institutionIllawarra Health & Medical Research Institute, Wollongong, New South Wales 2522, Australia
dc.contributor.institutionZernike Institute for Advanced Materials, University of Groningen, Groningen 9747 AG, The Netherlands
dc.contributor.institutionResearch School of Chemistry, Australian National University, Canberra, Australian Capital Territory 2601, Australia
dc.identifier.volume49
dc.identifier.issue12
dc.identifier.pages6804-6816
kaust.grant.numberOSR-2015-CRG4-2644
dc.identifier.eid2-s2.0-85109116714
dc.date.published-online2021-06-17
dc.date.published-print2021-07-09


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This is a pre-copyedited, author-produced PDF of an article accepted for publication in Nucleic acids research following peer review. The version of record is available online at: https://academic.oup.com/nar/article/49/12/6804/6303457.
Except where otherwise noted, this item's license is described as This is a pre-copyedited, author-produced PDF of an article accepted for publication in Nucleic acids research following peer review. The version of record is available online at: https://academic.oup.com/nar/article/49/12/6804/6303457.