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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:
Provided by: Nottingham ePrints

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  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.
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  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.
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  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.
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  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?
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  53. (2002). The structure of bacterial DnaA: implications for general mechanisms underlying DNA replication initiation.
  54. (2005). Unraveling the early steps of prokaryotic replication.
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