Interaction of the replication terminator protein of Bacillus subtilis with DNA probed by NMR spectroscopy
- School of Molecular and Microbial Biosciences, University of Sydney, Sydney, NSW 2006 (Australia)
- Research School of Chemistry, Australian National University, Canberra, ACT (Australia)
- Structural Chemistry Laboratory, AstraZeneca R and D, S-431 83, Moelndal (Sweden)
- Biochemistry and Molecular Biology, Monash University, Melbourne, Vic. 3800 (Australia)
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 in {sup 1}H-{sup 15}N correlation spectra of a {sup 15}N-labelled RTP.C110S mutant upon the addition of a 21 base pair symmetrical DNA binding site. Assignment of the {sup 1}H-{sup 15}N correlations was achieved using a suite of triple resonance NMR experiments with {sup 15}N,{sup 13}C,70% {sup 2}H enriched protein recorded at 800 MHz and using TROSY pulse sequences. Perturbations to {sup 1}H-{sup 15}N spectra revealed that the N-termini, {alpha}3-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.
- OSTI ID:
- 20710977
- Journal Information:
- Biochemical and Biophysical Research Communications, Vol. 335, Issue 2; Other Information: DOI: 10.1016/j.bbrc.2005.07.082; PII: S0006-291X(05)01544-5; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
Similar Records
Nucleotide sequence of Bacillus subtilis dnaB: a gene essential for DNA replication initiation and membrane attachment
Polypeptide backbone resonance assignments and secondary structure of Bacillus subtilis enzyme III sup glc determined by two-dimensional and three-dimensional heteronuclear NMR spectroscopy