1887

Abstract

A binding site for DnaA protein was identified in the regulatory region of the gene of . Specific binding was demonstrated by assays including filter binding as well as mobility shift in a polyacrylamide gel of the DnaA–DNA complex. In cells growing in minimal medium containing glucose, expression of β-galactosidase activity from an fusion gene was suppressed by oversupply of DnaA protein and was enhanced by reducing the free DnaA level. These results suggest that DnaA protein negatively regulates expression of the gene under these conditions. Despite fairly strong binding, the bound DNA fragment had no consensus 9 bp DnaA binding sequence (DnaA box), and anomalous binding to an AT-rich sequence located close to the transcription start site was revealed by a footprinting experiment.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-1-153
2001-01-01
2024-04-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/1/1470153a.html?itemId=/content/journal/micro/10.1099/00221287-147-1-153&mimeType=html&fmt=ahah

References

  1. Fuller, R. S., Funnell, B. E. & Kornberg, A.(1984). The DnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites. Cell 38, 889-900.[CrossRef] [Google Scholar]
  2. Hansen, F. G., Koefoed, S., Sørensen, L. & Atlung, T.(1987). Titration of DnaA protein by oriC DnaA-boxes increases dnaA gene expression in Escherichia coli. EMBO J 6, 255-258. [Google Scholar]
  3. Hidalgo, E., Chen, Y.-M., Lin, E. C. C. & Aguilar, J.(1991). Molecular cloning and DNA sequencing of the Escherichia coli K-12 ald gene encoding aldehyde dehydrogenase. J Bacteriol 173, 6118-6123. [Google Scholar]
  4. Kitagawa, R., Mitsuki, H., Okazaki, T. & Ogawa, T.(1996). A novel DnaA protein-binding site at 94·7 min on the Escherichia coli chromosome. Mol Microbiol 19, 1137-1147.[CrossRef] [Google Scholar]
  5. Kitagawa, R., Ozaki, T., Moriya, S. & Ogawa, T.(1998). Negative control of replication initiation by a novel chromosomal locus exhibiting exceptional affinity for Escherichia coli DnaA protein. Genes Dev 12, 3032-3043.[CrossRef] [Google Scholar]
  6. Kohara, Y., Akiyama, K. & Isono, K.(1987). The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell 50, 495-508.[CrossRef] [Google Scholar]
  7. Konopa, G., Szalewska-Palasz, A., Schmidt, A., Srutkowska, S., Messer, W. & Wegrzyn, G.(1999). The presence of two DnaA-binding sequences is required for an efficient interaction of the Escherichia coli DnaA protein with each particular weak DnaA box region. FEMS Microbiol Lett 174, 25-31.[CrossRef] [Google Scholar]
  8. Kornberg, A. & Baker, T. A. (1992).DNA Replication, 2nd edn. New York: Freeman.
  9. Limon, A., Hidalgo, E. & Aguilar, J.(1997). The aldA gene of Escherichia coli is under the control of at least three transcriptional regulators. Microbiology 143, 2085-2095.[CrossRef] [Google Scholar]
  10. Messer, W. & Weigel, C.(1997). DnaA initiator – also a transcription factor. Mol Microbiol 24, 1-6.[CrossRef] [Google Scholar]
  11. Miller, J. H. (1972).Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  12. Parada, C. A. & Marians, K. J.(1991). Mechanism of DnaA protein-dependent pBR322 DNA replication. DnaA protein-mediated trans-strand loading of the DnaB protein at the origin of pBR322 DNA. J Biol Chem 266, 18895-18906. [Google Scholar]
  13. Pellicer, M. T., Lynch, A. S., De Wulf, P., Boyd, D., Aguilar, J. & Lin, E. C. C.(1999). A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli. Mol Gen Genet 261, 170-176.[CrossRef] [Google Scholar]
  14. Quintilla, F. X., Baldoma, L., Badia, J. & Aguilar, J.(1991). Aldehyde dehydrogenase induction by glutamate in Escherichia coli. Role of 2-oxoglutarate. Eur J Biochem 202, 1321-1325.[CrossRef] [Google Scholar]
  15. Schaefer, C. & Messer, W.(1991). DnaA protein/DNA interaction. Modulation of the recognition sequence. Mol Gen Genet 226, 34-40. [Google Scholar]
  16. Schaper, S. & Messer, W.(1995). Interaction of the initiator protein DnaA of Escherichia coli with its DnaA target. J Biol Chem 270, 17622-17626.[CrossRef] [Google Scholar]
  17. Speck, C., Weigel, C. & Messer, W.(1999). ATP- and ADP-DnaA protein, a molecular switch in gene regulation. EMBO J 18, 6169-6176.[CrossRef] [Google Scholar]
  18. Straney, D. C., Straney, S. B. & Crothers, D. M.(1989). Synergy between Escherichia coli CAP protein and RNA polymerase in the lac promoter open complex. J Mol Biol 206, 41-57.[CrossRef] [Google Scholar]
  19. Yung, B. Y. & Kornberg, A.(1989). The DnaA initiator protein binds separate domains in the replication origin of Escherichia coli. J Biol Chem 264, 6146-6150. [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-1-153
Loading
/content/journal/micro/10.1099/00221287-147-1-153
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error