Handle URI:
http://hdl.handle.net/10754/596797
Title:
Loading dynamics of a sliding DNA clamp.
Authors:
Cho, Won-Ki; Jergic, Slobodan; Kim, Daehyung; Dixon, Nicholas E; Lee, Jong-Bong
Abstract:
Sliding DNA clamps are loaded at a ss/dsDNA junction by a clamp loader that depends on ATP binding for clamp opening. Sequential ATP hydrolysis results in closure of the clamp so that it completely encircles and diffuses on dsDNA. We followed events during loading of an E. coli β clamp in real time by using single-molecule FRET (smFRET). Three successive FRET states were retained for 0.3 s, 0.7 s, and 9 min: Hydrolysis of the first ATP molecule by the γ clamp loader resulted in closure of the clamp in 0.3 s, and after 0.7 s in the closed conformation, the clamp was released to diffuse on the dsDNA for at least 9 min. An additional single-molecule polarization study revealed that the interfacial domain of the clamp rotated in plane by approximately 8° during clamp closure. The single-molecule polarization and FRET studies thus revealed the real-time dynamics of the ATP-hydrolysis-dependent 3D conformational change of the β clamp during loading at a ss/dsDNA junction.
Citation:
Cho W-K, Jergic S, Kim D, Dixon NE, Lee J-B (2014) Loading Dynamics of a Sliding DNA Clamp. Angew Chem Int Ed 53: 6768–6771. Available: http://dx.doi.org/10.1002/anie.201403063.
Publisher:
Wiley-Blackwell
Journal:
Angewandte Chemie International Edition
Issue Date:
22-May-2014
DOI:
10.1002/anie.201403063
PubMed ID:
24854225
PubMed Central ID:
PMC4320747
Type:
Article
ISSN:
1433-7851
Sponsors:
This research was supported by the National Research Foundation (NRF) of Korea and was funded by the Ministry of Education, Science, and Technology (MEST; Grants No. 2011-0013901 and No. 2010-0019706), by the Australian Research Council (DP0877658), and by an FIC Award from the King Abdullah University of Science and Technology (Saudi Arabia).
Appears in Collections:
Publications Acknowledging KAUST Support

Full metadata record

DC FieldValue Language
dc.contributor.authorCho, Won-Kien
dc.contributor.authorJergic, Slobodanen
dc.contributor.authorKim, Daehyungen
dc.contributor.authorDixon, Nicholas Een
dc.contributor.authorLee, Jong-Bongen
dc.date.accessioned2016-02-21T08:50:52Zen
dc.date.available2016-02-21T08:50:52Zen
dc.date.issued2014-05-22en
dc.identifier.citationCho W-K, Jergic S, Kim D, Dixon NE, Lee J-B (2014) Loading Dynamics of a Sliding DNA Clamp. Angew Chem Int Ed 53: 6768–6771. Available: http://dx.doi.org/10.1002/anie.201403063.en
dc.identifier.issn1433-7851en
dc.identifier.pmid24854225en
dc.identifier.doi10.1002/anie.201403063en
dc.identifier.urihttp://hdl.handle.net/10754/596797en
dc.description.abstractSliding DNA clamps are loaded at a ss/dsDNA junction by a clamp loader that depends on ATP binding for clamp opening. Sequential ATP hydrolysis results in closure of the clamp so that it completely encircles and diffuses on dsDNA. We followed events during loading of an E. coli β clamp in real time by using single-molecule FRET (smFRET). Three successive FRET states were retained for 0.3 s, 0.7 s, and 9 min: Hydrolysis of the first ATP molecule by the γ clamp loader resulted in closure of the clamp in 0.3 s, and after 0.7 s in the closed conformation, the clamp was released to diffuse on the dsDNA for at least 9 min. An additional single-molecule polarization study revealed that the interfacial domain of the clamp rotated in plane by approximately 8° during clamp closure. The single-molecule polarization and FRET studies thus revealed the real-time dynamics of the ATP-hydrolysis-dependent 3D conformational change of the β clamp during loading at a ss/dsDNA junction.en
dc.description.sponsorshipThis research was supported by the National Research Foundation (NRF) of Korea and was funded by the Ministry of Education, Science, and Technology (MEST; Grants No. 2011-0013901 and No. 2010-0019706), by the Australian Research Council (DP0877658), and by an FIC Award from the King Abdullah University of Science and Technology (Saudi Arabia).en
dc.publisherWiley-Blackwellen
dc.rightsThis is an open access article under the terms of the Creative Commons Attribution Non-Commercial NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectDNA replicationen
dc.subjectTernary Complexesen
dc.subjectSingle-molecule Freten
dc.subjectSingle-molecule Polarizationen
dc.subjectDna Clampsen
dc.titleLoading dynamics of a sliding DNA clamp.en
dc.typeArticleen
dc.identifier.journalAngewandte Chemie International Editionen
dc.identifier.pmcidPMC4320747en
dc.contributor.institutionDepartment of Physics, School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology (POSTECH), Pohang (Korea).en

Related articles on PubMed

All Items in KAUST are protected by copyright, with all rights reserved, unless otherwise indicated.