A direct proofreader-clamp interaction stabilizes the Pol III replicase in the polymerization mode

Handle URI:
http://hdl.handle.net/10754/562660
Title:
A direct proofreader-clamp interaction stabilizes the Pol III replicase in the polymerization mode
Authors:
Jergic, Slobodan; Horan, Nicholas P.; Elshenawy, Mohamed ( 0000-0002-8599-8388 ) ; Mason, Claire E.; Urathamakul, Thitima; Ozawa, Kiyoshi; Robinson, Andrew J.; Goudsmits, Joris M H; Wang, Yao; Pan, Xuefeng; Beck, Jennifer L.; Van Oijen, Antoine M.; Huber, Thomas L.; Hamdan, Samir ( 0000-0001-5192-1852 ) ; Dixon, Nicholas E.
Abstract:
Processive DNA synthesis by the αÉ"θ core of the Escherichia coli Pol III replicase requires it to be bound to the β 2 clamp via a site in the α polymerase subunit. How the É" proofreading exonuclease subunit influences DNA synthesis by α was not previously understood. In this work, bulk assays of DNA replication were used to uncover a non-proofreading activity of É". Combination of mutagenesis with biophysical studies and single-molecule leading-strand replication assays traced this activity to a novel β-binding site in É" that, in conjunction with the site in α, maintains a closed state of the αÉ"θ-β 2 replicase in the polymerization mode of DNA synthesis. The É"-β interaction, selected during evolution to be weak and thus suited for transient disruption to enable access of alternate polymerases and other clamp binding proteins, therefore makes an important contribution to the network of protein-protein interactions that finely tune stability of the replicase on the DNA template in its various conformational states. © 2013 European Molecular Biology Organization.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division; Bioscience Program
Publisher:
EMBO Press
Journal:
EMBO Journal
Issue Date:
22-Feb-2013
DOI:
10.1038/emboj.2012.347
PubMed ID:
23435564
PubMed Central ID:
PMC3642676
Type:
Article
ISSN:
02614189
Sponsors:
We thank Michelle Blayney and Linda Jessop for preliminary ESI-MS data. This work was supported by grants from the Australian Research Council, including Fellowships to KO, TH, and NED, and by a KAUST Faculty Initiated Collaborative grant to SMH and NED.
Additional Links:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642676
Appears in Collections:
Articles; Bioscience Program; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorJergic, Slobodanen
dc.contributor.authorHoran, Nicholas P.en
dc.contributor.authorElshenawy, Mohameden
dc.contributor.authorMason, Claire E.en
dc.contributor.authorUrathamakul, Thitimaen
dc.contributor.authorOzawa, Kiyoshien
dc.contributor.authorRobinson, Andrew J.en
dc.contributor.authorGoudsmits, Joris M Hen
dc.contributor.authorWang, Yaoen
dc.contributor.authorPan, Xuefengen
dc.contributor.authorBeck, Jennifer L.en
dc.contributor.authorVan Oijen, Antoine M.en
dc.contributor.authorHuber, Thomas L.en
dc.contributor.authorHamdan, Samiren
dc.contributor.authorDixon, Nicholas E.en
dc.date.accessioned2015-08-03T11:00:13Zen
dc.date.available2015-08-03T11:00:13Zen
dc.date.issued2013-02-22en
dc.identifier.issn02614189en
dc.identifier.pmid23435564en
dc.identifier.doi10.1038/emboj.2012.347en
dc.identifier.urihttp://hdl.handle.net/10754/562660en
dc.description.abstractProcessive DNA synthesis by the αÉ"θ core of the Escherichia coli Pol III replicase requires it to be bound to the β 2 clamp via a site in the α polymerase subunit. How the É" proofreading exonuclease subunit influences DNA synthesis by α was not previously understood. In this work, bulk assays of DNA replication were used to uncover a non-proofreading activity of É". Combination of mutagenesis with biophysical studies and single-molecule leading-strand replication assays traced this activity to a novel β-binding site in É" that, in conjunction with the site in α, maintains a closed state of the αÉ"θ-β 2 replicase in the polymerization mode of DNA synthesis. The É"-β interaction, selected during evolution to be weak and thus suited for transient disruption to enable access of alternate polymerases and other clamp binding proteins, therefore makes an important contribution to the network of protein-protein interactions that finely tune stability of the replicase on the DNA template in its various conformational states. © 2013 European Molecular Biology Organization.en
dc.description.sponsorshipWe thank Michelle Blayney and Linda Jessop for preliminary ESI-MS data. This work was supported by grants from the Australian Research Council, including Fellowships to KO, TH, and NED, and by a KAUST Faculty Initiated Collaborative grant to SMH and NED.en
dc.publisherEMBO Pressen
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3642676en
dc.subjectbeta sliding clampen
dc.subjectDNA polymerase IIIen
dc.subjectDNA replicationen
dc.subjectEscherichia colien
dc.subjectproofreading exonucleaseen
dc.titleA direct proofreader-clamp interaction stabilizes the Pol III replicase in the polymerization modeen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentBioscience Programen
dc.identifier.journalEMBO Journalen
dc.identifier.pmcidPMC3642676en
dc.contributor.institutionSchool of Chemistry, University of Wollongong, Northfields Avenue, Wollongong, NSW 2522, Australiaen
dc.contributor.institutionResearch School of Chemistry, Australian National University, Canberra, ACT, Australiaen
dc.contributor.institutionZernike Institute for Advanced Materials, Groningen, Netherlandsen
dc.contributor.institutionDepartment of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, United Statesen
dc.contributor.institutionSchool of Life Science, Beijing Institute of Technology, Beijing 100081, Chinaen
kaust.authorHamdan, Samiren
kaust.authorElshenawy, Mohameden

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