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 Ogura, Y.
Right arrow Articles by Moriya, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ogura, Y.
Right arrow Articles by Moriya, S.

 Previous Article  |  Next Article 

Journal of Bacteriology, July 2001, p. 3833-3841, Vol. 183, No. 13
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.13.3833-3841.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Autoregulation of the dnaA-dnaN Operon and Effects of DnaA Protein Levels on Replication Initiation in Bacillus subtilis

Yoshitoshi Ogura, Yukiho Imai,dagger Naotake Ogasawara, and Shigeki Moriya*

Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630-0101, Japan

Received 18 December 2000/Accepted 6 April 2001

In Escherichia coli, the DnaA protein level appears to play a pivotal role in determining the timing of replication initiation. To examine the effects on replication initiation in B. subtilis, we constructed a strain in which a copy of the dnaA gene was integrated at the purA locus on the chromosome under the control of an isopropyl-beta -D-thiogalactopyranoside (IPTG)-inducible promoter. However, increasing the DnaA level resulted in cell elongation and inhibition of cell growth by induction of the SOS response. Transcription of the native dnaA-dnaN operon was greatly reduced at high DnaA levels, but it was increased in a dnaA-null mutant, indicating autoregulation of the operon by DnaA. When a copy of the dnaN gene was added downstream of the additional dnaA gene at purA, the cells grew at high DnaA levels, suggesting that depletion of DnaN (beta  subunit of DNA polymerase III) within the cell by repression of the native dnaA-dnaN operon at high DnaA levels was the cause of the SOS induction. Flow cytometry of the cells revealed that the cell mass at initiation of replication increased at a lower DnaA level and decreased at DnaA levels higher than those of the wild type. Proper timing of replication initiation was observed at DnaA levels nearly comparable to the wild-type level. These results suggest that if the DnaA level increases with progression of the replication cycle, it could act as a rate-limiting factor of replication initiation in B. subtilis.

* Corresponding author. Mailing address: Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5, Takayama, Ikoma, Nara 630-0101, Japan. Phone: 81-743-72-5432. Fax: 81-743-72-5439. E-mail: moriya{at}bs.aist-nara.ac.jp.

dagger Present address: Genox Research, Inc., Kawasaki Laboratory, Teikyo Univ. Biotech. Center, 907 Nogawa, Miyamae-ku, Kawasaki, Kanagawa 216-0001, Japan.

Journal of Bacteriology, July 2001, p. 3833-3841, Vol. 183, No. 13
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.13.3833-3841.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Breier, A. M., Grossman, A. D. (2009). Dynamic Association of the Replication Initiator and Transcription Factor DnaA with the Bacillus subtilis Chromosome during Replication Stress. J. Bacteriol. 191: 486-493 [Abstract] [Full Text]  
  • Ishikawa, S., Ogura, Y., Yoshimura, M., Okumura, H., Cho, E., Kawai, Y., Kurokawa, K., Oshima, T., Ogasawara, N. (2007). Distribution of Stable DnaA-Binding Sites on the Bacillus Subtilis Genome Detected using a Modified ChIP-chip Method. DNA Res 0: dsm017v1-14 [Abstract] [Full Text]  
  • Simmons, L. A., Grossman, A. D., Walker, G. C. (2007). Replication is required for the RecA localization response to DNA damage in Bacillus subtilis. Proc. Natl. Acad. Sci. USA 104: 1360-1365 [Abstract] [Full Text]  
  • Noirot-Gros, M.-F.ço., Velten, M., Yoshimura, M., McGovern, S., Morimoto, T., Ehrlich, S. D., Ogasawara, N., Polard, P., Noirot, P. (2006). Functional dissection of YabA, a negative regulator of DNA replication initiation in Bacillus subtilis. Proc. Natl. Acad. Sci. USA 103: 2368-2373 [Abstract] [Full Text]  
  • Goranov, A. I., Katz, L., Breier, A. M., Burge, C. B., Grossman, A. D. (2005). A transcriptional response to replication status mediated by the conserved bacterial replication protein DnaA. Proc. Natl. Acad. Sci. USA 102: 12932-12937 [Abstract] [Full Text]  
  • Capaldi, S. A., Berger, J. M. (2004). Biochemical characterization of Cdc6/Orc1 binding to the replication origin of the euryarchaeon Methanothermobacter thermoautotrophicus. Nucleic Acids Res 32: 4821-4832 [Abstract] [Full Text]  
  • Wang, L., Trawick, J. D., Yamamoto, R., Zamudio, C. (2004). Genome-wide operon prediction in Staphylococcus aureus. Nucleic Acids Res 32: 3689-3702 [Abstract] [Full Text]  
  • Ogura, Y., Ogasawara, N., Harry, E. J., Moriya, S. (2003). Increasing the Ratio of Soj to Spo0J Promotes Replication Initiation in Bacillus subtilis. J. Bacteriol. 185: 6316-6324 [Abstract] [Full Text]  
  • Salazar, L., Guerrero, E., Casart, Y., Turcios, L., Bartoli, F. (2003). Transcription analysis of the dnaA gene and oriC region of the chromosome of Mycobacterium smegmatis and Mycobacterium bovis BCG, and its regulation by the DnaA protein. Microbiology 149: 773-784 [Abstract] [Full Text]  
  • Lee, P. S., Lin, D. C.-H., Moriya, S., Grossman, A. D. (2003). Effects of the Chromosome Partitioning Protein Spo0J (ParB) on oriC Positioning and Replication Initiation in Bacillus subtilis. J. Bacteriol. 185: 1326-1337 [Abstract] [Full Text]  
  • Grigorian, A. V., Lustig, R. B., Guzman, E. C., Mahaffy, J. M., Zyskind, J. W. (2003). Escherichia coli Cells with Increased Levels of DnaA and Deficient in Recombinational Repair Have Decreased Viability. J. Bacteriol. 185: 630-644 [Abstract] [Full Text]  
  • Greendyke, R., Rajagopalan, M., Parish, T., Madiraju, M. V. V. S. (2002). Conditional expression of Mycobacterium smegmatis dnaA, an essential DNA replication gene. Microbiology 148: 3887-3900 [Abstract] [Full Text]  
  • Morimoto, T., Loh, P. C., Hirai, T., Asai, K., Kobayashi, K., Moriya, S., Ogasawara, N. (2002). Six GTP-binding proteins of the Era/Obg family are essential for cell growth in Bacillus subtilis. Microbiology 148: 3539-3552 [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»