This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kurz, M.
Right arrow Articles by Kongsuwan, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kurz, M.
Right arrow Articles by Kongsuwan, K.

 Previous Article  |  Next Article 

Journal of Bacteriology, June 2004, p. 3508-3515, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3508-3515.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Interaction of the Sliding Clamp ß-Subunit and Hda, a DnaA-Related Protein

Mareike Kurz,{dagger} Brian Dalrymple, Gene Wijffels, and Kritaya Kongsuwan*

CSIRO Livestock Industries, Queensland Bioscience Precinct, St. Lucia Queensland Dominion 4067, Australia

Received 23 October 2003/ Accepted 12 February 2004

In Escherichia coli, interactions between the replication initiation protein DnaA, the ß subunit of DNA polymerase III (the sliding clamp protein), and Hda, the recently identified DnaA-related protein, are required to convert the active ATP-bound form of DnaA to an inactive ADP-bound form through the accelerated hydrolysis of ATP. This rapid hydrolysis of ATP is proposed to be the main mechanism that blocks multiple initiations during cell cycle and acts as a molecular switch from initiation to replication. However, the biochemical mechanism for this crucial step in DNA synthesis has not been resolved. Using purified Hda and ß proteins in a plate binding assay and Ni-nitrilotriacetic acid pulldown analysis, we show for the first time that Hda directly interacts with ß in vitro. A new ß-binding motif, a hexapeptide with the consensus sequence QL[SP]LPL, related to the previously identified ß-binding pentapeptide motif (QL[SD]LF) was found in the amino terminus of the Hda protein. Mutants of Hda with amino acid changes in the hexapeptide motif are severely defective in their ability to bind ß. A 10-amino-acid peptide containing the E. coli Hda ß-binding motif was shown to compete with Hda for binding to ß in an Hda-ß interaction assay. These results establish that the interaction of Hda with ß is mediated through the hexapeptide sequence. We propose that this interaction may be crucial to the events that lead to the inactivation of DnaA and the prevention of excess initiation of rounds of replication.

* Corresponding author: Mailing address: CSIRO Livestock Industries, Queensland Bioscience Precinct, 306 Carmody Rd., St. Lucia QLD 4067, Australia. Phone: 617 3214 2512. Fax: 61 7 3214 2509. E-mail: kritaya.kongsuwan{at}csiro.au.

{dagger} Present address: Institute of Molecular Biosciences, University of Queensland, St. Lucia QLD 4067, Australia.

Journal of Bacteriology, June 2004, p. 3508-3515, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3508-3515.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Su'etsugu, M., Nakamura, K., Keyamura, K., Kudo, Y., Katayama, T. (2008). Hda Monomerization by ADP Binding Promotes Replicase Clamp-mediated DnaA-ATP Hydrolysis. J. Biol. Chem. 283: 36118-36131 [Abstract] [Full Text]  
  • Kongsuwan, K., Josh, P., Picault, M. J., Wijffels, G., Dalrymple, B. (2006). The Plasmid RK2 Replication Initiator Protein (TrfA) Binds to the Sliding Clamp {beta} Subunit of DNA Polymerase III: Implication for the Toxicity of a Peptide Derived from the Amino-Terminal Portion of 33-Kilodalton TrfA.. J. Bacteriol. 188: 5501-5509 [Abstract] [Full Text]  
  • Banack, T., Clauson, N., Ogbaa, N., Villar, J., Oliver, D., Firshein, W. (2005). Overexpression of the Hda DnaA-Related Protein in Escherichia coli Inhibits Multiplication, Affects Membrane Permeability, and Induces the SOS Response. J. Bacteriol. 187: 8507-8510 [Full Text]  
  • Su'etsugu, M., Shimuta, T.-r., Ishida, T., Kawakami, H., Katayama, T. (2005). Protein Associations in DnaA-ATP Hydrolysis Mediated by the Hda-Replicase Clamp Complex. J. Biol. Chem. 280: 6528-6536 [Abstract] [Full Text]  
  • Sutton, M. D. (2004). The Escherichia coli dnaN159 Mutant Displays Altered DNA Polymerase Usage and Chronic SOS Induction. J. Bacteriol. 186: 6738-6748 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»