Location of Repository

DnaB proteolysis in vivo regulates oligomerization and its localization at oriC in Bacillus subtilis

By William H. Grainger, Cristina Machón, David J. Scott and Panos Soultanas


Initiation of bacterial DNA replication at oriC is mediated by primosomal proteins that act cooperatively to melt an AT-rich region where the replicative helicase is loaded prior to the assembly of the replication fork. In Bacillus subtilis, the dnaD, dnaB and dnaI genes are essential for initiation of DNA replication. We established that their mRNAs are maintained in fast growing asynchronous cultures. DnaB is truncated at its C-terminus in a growth phase-dependent manner. Proteolysis is confined to cytosolic, not to membrane-associated DnaB, and affects oligomerization. Truncated DnaB is depleted at the oriC relative to the native protein. We propose that DNA-induced oligomerization is essential for its action at oriC and proteolysis regulates its localization at oriC. We show that DnaB has two separate ssDNA-binding sites one located within residues 1-300 and another between residues 365-428, and a dsDNA-binding site within residues 365-428. Tetramerization of DnaB is mediated within residues 1-300, and DNA-dependent oligomerization within residues 365-428. Finally, we show that association of DnaB with the oriC is asymmetric and extensive. It encompasses an area from the middle of dnaA to the end of yaaA that includes the AT-rich region melted during the initiation stage of DNA replication

Publisher: Oxford Journals
Year: 2010
OAI identifier: oai:eprints.nottingham.ac.uk:1311
Provided by: Nottingham ePrints

Suggested articles



  1. (2002). A functional interaction between the putative primosomal protein DnaI and the main replicative DNA helicase DnaB in Bacillus.
  2. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
  3. (1991). A sequence similarity between proteins involved in initiation and termination of bacterial chromosome replication.
  4. (2005). A truncated H-NS-like protein from enteropathogenic Escherichia coli acts as an H-NS antagonist. doi
  5. (2003). A two-protein strategy for the functional loading of a cellular replicative DNA helicase.
  6. (1992). Amino acid substitution matrices from protein blocks.
  7. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2CT method.
  8. (2001). Autoregulation of the dnaA-dnaN operon and effects of DnaA protein levels on replication initiation in Bacillus subtilis.
  9. (1998). Cell membrane and chromosome replication in Bacillus subtilis.
  10. (2007). Clustal W and Clustal X version
  11. (1997). Complexes at the replication origin of Bacillus subtilis with homologous and heterologous DnaA protein. doi
  12. (2004). Control of DNA replication initiation by recruitment of an essential initiation protein to the membrane of Bacillus subtilis.
  13. (2007). DNA replication initiation: mechanisms and regulation in bacteria.
  14. (1980). DNA-membrane association is necessary for initiation of chromosomal and plasmid replication in Bacillus subtilis.
  15. (2006). DnaA: controlling the initiation of bacterial DNA replication and more.
  16. (2007). DnaB and DnaI temperature-sensitive mutants of Staphylococcus aureus: evidence for involvement of DnaB and DnaI in synchrony regulation of chromosome replication.
  17. (2001). DnaB, DnaD and DnaI proteins are components of the Bacillus subtilis replication restart primosome.
  18. (2001). DnaD protein of Bacillus subtilis interacts with DnaA, the initiator protein of replication.
  19. (1999). Domain organization and oligomerisation among H-NS-like nucleoidassociated proteins in bacteria.
  20. (2001). Early steps of Bacillus subtilis primosome assembly.
  21. (1987). Effects of temperature sensitive variants of the Bacillus subtilis dnaB gene on the replication of low-copy-number plasmid.
  22. (2005). Functional interplay between the Bacillus subtilis DnaD and DnaB proteins essential for initiation and re-initiation of DNA replication.
  23. (2008). Genome-wide analysis of Fis binding in Escherichia coli indicates a causative role for A-/AT-tracts.
  24. (2004). Growth and sporulation of Bacillus cereus ATCC 14579 under defined conditions: temporal expression of genes for key sigma factors.
  25. (2006). Helicase binding to DnaI exposes a cryptic DNA-binding site during helicase loading in Bacillus subtilis. doi
  26. (2004). Identification of temperature-sensitive dnaD mutants of Staphylococcus aureus that are defective in chromosome DNA replication.
  27. (1987). Identification of the product of dnaB gene in Bacillus subtilis.
  28. (1991). In vitro type II binding of chromosomal DNA to membrane in Bacillus subtilis.
  29. (2007). Loading a ring: Structure of the Bacillus subtilis DnaB protein, a co-loader of the replicative helicase.
  30. (1986). Nucleotide sequence and organization of dnaB gene and neighbouring geneson the Bacillus subtilis chromosome.
  31. (1987). Nucleotide sequence of Bacillus subtilis dnaB: a gene essential for DNA replication initiation and membrane attachment.
  32. (2003). On the analysis of protein self-association by sedimentation velocity analytical ultracentrifugation.
  33. (1995). Primosome assembly site in Bacillus subtilis.
  34. (1996). Probing the structure function and interactions of the Escherichia coli H-NS Nucleic Acids Research,
  35. (2006). Replisome assembly and the direct restart of stalled replication forks.
  36. (2001). Requirement for the molecular adapter function of StpA at the Escherichia coli bgl promoter depends upon the level of truncated H-NS protein. doi
  37. (1998). Sedimentation analysis of noninteracting and self-associating solutes using numerical solutions to the Lamm equation.
  38. (2008). Single-molecule atomic force spectroscopy reveals that DnaD forms scaffolds and enhances duplex melting.
  39. (2000). Size-distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and Lamm equation modeling.
  40. (2008). Structural synergy and molecular crosstalk between bacterial helicase loaders and replication initiators.
  41. (2008). Structure of the N-terminal oligomerisation domain of DnaD reveals a unique tetramerization motif and provides insights into scaffold formation.
  42. (2006). Systematic localization of proteins fused to the green fluorescent protein in Bacillus subtilis: identification of new proteins at the DNA replication factory.
  43. (1977). Technical considerations on the use of horseradish peroxidase as a neuronal marker.
  44. (2005). The Bacillus subtilis DnaD and DnaB proteins exhibit different DNA remodeling activities.
  45. (2004). The Bacillus subtilis DnaD protein: a putative link between DNA remodeling and initiation of DNA replication.
  46. (1995). The Bacillus subtilis dnaI gene is part of the dnaB operon.
  47. (2006). The Bacillus subtilis primosomal protein DnaD untwists supercoiled DNA.
  48. (2002). The bacterial replication initiator DnaA. DnaA and oriC, the bacterial mode to initiate DNA replication.
  49. (2005). The disposition of nascent strands at stalled replication forks dictates the pathway of replisome loading during restart.
  50. (2006). The DNA-remodelling activity of DnaD is the sum of oligomerisation and DNA-binding activities on separate domains.
  51. (1996). The initiation mess?
  52. (2008). The Jpred 3 secondaray structure prediction server.
  53. (2002). The structure of bacterial DnaA: implications for general mechanisms underlying DNA replication initiation.
  54. (2005). Unraveling the early steps of prokaryotic replication.
  55. (2004). Where does bacterial replication start? Rules for predicting the oriC region.

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.