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Interaction of the replication terminator protein of Bacillus subtilis with DNA probed by NMR spectroscopy

Hastings, Adam; Folmer, Rutger; Duggin, Iain; Wake, R. Gerry; Wilce, Matthew; Wilce, Jacqueline; Otting, Gottfried

Description

Termination of DNA replication in Bacillus subtilis involves the polar arrest of replication forks by a specific complex formed between the dimeric 29 kDa replication terminator protein (RTP) and DNA terminator sites. We have used NMR spectroscopy to probe the changes in1H-15N correlation spectra of a15N-labelled RTP.C110S mutant upon the addition of a 21 base pair symmetrical DNA binding site. Assignment of the1H-15N correlations was achieved using a suite of triple resonance NMR experiments...[Show more]

dc.contributor.authorHastings, Adam
dc.contributor.authorFolmer, Rutger
dc.contributor.authorDuggin, Iain
dc.contributor.authorWake, R. Gerry
dc.contributor.authorWilce, Matthew
dc.contributor.authorWilce, Jacqueline
dc.contributor.authorOtting, Gottfried
dc.date.accessioned2015-12-13T22:53:51Z
dc.identifier.issn0006-291X
dc.identifier.urihttp://hdl.handle.net/1885/81995
dc.description.abstractTermination of DNA replication in Bacillus subtilis involves the polar arrest of replication forks by a specific complex formed between the dimeric 29 kDa replication terminator protein (RTP) and DNA terminator sites. We have used NMR spectroscopy to probe the changes in1H-15N correlation spectra of a15N-labelled RTP.C110S mutant upon the addition of a 21 base pair symmetrical DNA binding site. Assignment of the1H-15N correlations was achieved using a suite of triple resonance NMR experiments with15N,13C,70%2H enriched protein recorded at 800 MHz and using TROSY pulse sequences. Perturbations to1H-15N spectra revealed that the N-termini, α3-helices and several loops are affected by the binding interaction. An analysis of this data in light of the crystallographically determined apo- and DNA-bound forms of RTP.C110S revealed that the NMR spectral perturbations correlate more closely to protein structural changes upon complex formation rather than to interactions at the protein-DNA interface.
dc.publisherAcademic Press
dc.sourceBiochemical and Biophysical Research Communications
dc.subjectKeywords: DNA; amino terminal sequence; article; Bacillus subtilis; binding site; correlation analysis; crystallography; DNA binding; DNA replication; nonhuman; nuclear magnetic resonance spectroscopy; priority journal; protein binding; protein interaction; Bacillu Duplex DNA; HSQC titration; NMR spectroscopy; Protein-DNA complex; RTP; Structural perturbation; Terminator DNA; TROSY; Winged-helix protein
dc.titleInteraction of the replication terminator protein of Bacillus subtilis with DNA probed by NMR spectroscopy
dc.typeJournal article
local.description.notesImported from ARIES
local.description.refereedYes
local.identifier.citationvolume335
dc.date.issued2005
local.identifier.absfor069999 - Biological Sciences not elsewhere classified
local.identifier.ariespublicationMigratedxPub10298
local.type.statusPublished Version
local.contributor.affiliationHastings, Adam, University of Sydney
local.contributor.affiliationOtting, Gottfried, College of Physical and Mathematical Sciences, ANU
local.contributor.affiliationFolmer, Rutger, Structural Chemistry Laboratory
local.contributor.affiliationDuggin, Iain, University of Sydney
local.contributor.affiliationWake, R. Gerry, University of Sydney
local.contributor.affiliationWilce, Matthew, University of Western Australia
local.contributor.affiliationWilce, Jacqueline, Monash University
local.description.embargo2037-12-31
local.bibliographicCitation.issue2
local.bibliographicCitation.startpage361
local.bibliographicCitation.lastpage366
local.identifier.doi10.1016/j.bbrc.2005.07.082
dc.date.updated2015-12-11T11:00:11Z
local.identifier.scopusID2-s2.0-23744473897
CollectionsANU Research Publications

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