Interaction of the replication terminator protein of Bacillus subtilis with DNA probed by NMR spectroscopy
Date
2005
Authors
Hastings, Adam
Folmer, Rutger
Duggin, Iain
Wake, R. Gerry
Wilce, Matthew
Wilce, Jacqueline
Otting, Gottfried
Journal Title
Journal ISSN
Volume Title
Publisher
Academic Press
Abstract
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 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.
Description
Keywords
Keywords: 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
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Source
Biochemical and Biophysical Research Communications
Type
Journal article
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2037-12-31