Skip to main content
Log in

DnaA protein/DNA interaction. Modulation of the recognition sequence

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

In the active orientation the DnaA protein/dnaA box complex blocks transcribing RNA polymerase. The extent of transcription termination at different dnaA boxes was used to determine in vivo their various affinities to the DnaA protein. The rate of transcription distal to the respective dnaA box was monitored by the expression of the reporter gene galK. The dnaA boxes (5′-TTATACACA and 5′-TTATCCAAA), present in oriC, showed the strongest binding affinity. The dnaA box 5′-TTTTCCACA was mutated at eight positions such that the boxes differed from the consensus sequence, 5′-TT(A/T)T(A/C)CA(A/C)A, defined so far. Based on the different properties of the dnaA boxes, a new consensus sequence was derived: 5′-(T/C)(T/C)(A/T/C)T(A/C)C(A/G)(A/C/T)(A/C).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Abeles AL, Reaves LD, Austin SJ (1990) A single dnaA box is sufficient for initiation from the P1 plasmid origin. J Bacteriol 172:4386–4391

    Google Scholar 

  • Atlung T, Clausen E, Hansen FG (1985) Autoregulation of the dnaA gene of Escherichia coli. Mol Gen Genet 200:442–450

    Google Scholar 

  • Bachmann BJ (1983) Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev 47:180–230

    Google Scholar 

  • Backman K, Ptashne M (1978) Maximizing gene expression on a plasmid using recombination in vitro. Cell 13:65–71

    Google Scholar 

  • Braun RE, O'Day K, Wright A (1985) Autoregulation of the DNA replication gene dnaA in E. coli. Cell 40:159–169

    Google Scholar 

  • Chiaramello AE, Zyskind JW (1989) Expression of Escherichia coli dnaA and mioC genes as a function of growth rate. J Bacteriol 171:4272–4280

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    Google Scholar 

  • Fuerste JP, Pansegrau W, Frank R, Bloecker H, Scholz P, Bagdasarian M, Lanka E (1986) Molecular cloning of the plasmid RP4 primase region in a multi-host range tacP expression vector. Gene 48:117–129

    Google Scholar 

  • Fukuoka T, Moriya S, Yoshikawa H, Ogasawara N (1990) Purification and characterization of an initiation protein for chromosomal replication, DnaA, in Bacillus subtilis. J Biochem 107:732–739

    Google Scholar 

  • Fuller RS, Funnell BE, Kornberg A (1984) The dnaA protein complex with the E. coli chromosomal origin (oriC) and other sites. Cell 38:889–900

    Google Scholar 

  • Gielow A, Kücherer C, Kölling R, Messer W (1988) Transcription in the region of the replication origin, oriC, of Escherichia coli. Termination of asnC transcripts. Mol Gen Genet 214:474–481

    Google Scholar 

  • Hansen FG, Koefoed S, Soerensen 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 

  • Jonczyk P, Hines R, Smith DW (1989) The Escherichia coli dam gene is expressed as a distal gene of a new operon. Mol Gen Genet 217:85–96

    Google Scholar 

  • Kücherer C, Lother H, Kölling R, Schauzu MA, Messer W (1986) Regulation of transcription of the chromosomal dnaA gene of Escherichia coli. Mol Gen Genet 205:115–121

    Google Scholar 

  • Loebner-Olesen A, Atlung T, Rasmussen KV (1987) Stability and replication control of E. coli minichromosomes. J Bacteriol 169:2835–2842

    Google Scholar 

  • Lother H, Kölling R, Kücherer C, Schauzu M (1985) Initiation of replication in Escherichia coli involves dnaA protein regulated transcription within the replication origin. EMBO J 4:555–560

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Masai H, Arai K-I (1987) RepA and DnaA proteins are required for initiation of R1 plasmid replication in vitro and interact with the oriR sequence. Proc Natl Acad Sci USA 84:4781–4785

    Google Scholar 

  • Matsui M, Oka A, Takanami M, Yasuda S, Hirota Y (1985) Sites of dnaA protein-binding in the replication origin of the E. coli K-12 chromosome. J Mol Biol 184:529–533

    Google Scholar 

  • McKenney K, Shimatake H, Court D, Schmeissner U, Brady C, Rosenberg M (1981) A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. In: Chirikijan JG, Papas TS (eds) Gene amplification and analysis. 2: Analysis of nucleic acids by enzymatic methods. North Holland Publishing Company, Amsterdam, pp 383–415

    Google Scholar 

  • Messer W, Seufert W, Schaefer C, Gielow A, Hartmann H, Wende M (1988) Functions of the DnaA protein of Escherichia coli in replication and transcription. Biochim Biophys Acta 951:351–358

    Google Scholar 

  • Moriya S, Fukuoka T, Ogasawara N, Yoshikawa H (1988) Regulation of initiation of the chromosomal replication by DnaA-boxes in the origin of the Bacillus subtilis chromosome. EMBO J 7:2911–2917

    Google Scholar 

  • Ortega S, Lanka E, Diaz R (1986) The involvement of host replication proteins and of specific origin sequences in the in vitro replication of miniplasmid R1 DNA. Nucleic Acids Res 14:4865–4879

    Google Scholar 

  • Polaczek P, Ciesla Z (1984) Effect of altered efficiency of the RNA I and RNA II promoters on in vivo replication of co1E1-like plasmids in Escherichia coli. Mol Gen Genet 194:227–231

    Google Scholar 

  • Rokeach LA, Chiaramello A, Junker DE, Crain K, Nourani A, Jannatipour M, Zyskind JW (1986) Effects of DnaA protein on replication and transcription events at the Escherichia coli origin of replication, oriC. UCLA Symp Mol Cell Biol 47:415–427

    Google Scholar 

  • Samitt CE, Hansen FG, Miller JF, Schaechter M (1989) In vivo studies of DnaA binding to the origin of replication of Escherichia coli. EMBO J 8:989–993

    Google Scholar 

  • Sargent MG (1968) Rapid fixed-time assay for penicillinase. J Bacteriol 95:1493–1494

    Google Scholar 

  • Schaefer C, Messer W (1988) Termination of the E. coli asnC transcript. The DnaA protein/dnaA box complex blocks transcribing RNA polymerase. Gene 73:347–354

    Google Scholar 

  • Schaefer C, Messer W (1989) Directionality of DnaA protein/DNA interaction. Active orientation of the DnaA protein/dnaA box complex in transcription termination. EMBO J 8:1609–1613

    Google Scholar 

  • Schauzu MA, Kücherer C, Kölling R, Messer W, Lother H (1987) Transcripts within the replication origin, oriC. of Escherichia coli. Nucleic Acids Res 15:2479–2497

    Google Scholar 

  • Sekimizu K, Bramhill D, Kornberg A (1988a) Sequential early stages in the in vitro initiation of replication at the origin of the Escherichia coli chromosome. J Biol Chem 263:7124–7130

    Google Scholar 

  • Sekimizu K, Yung BY, Kornberg A (1988b) The dnaA protein of Escherichia coli. Abundance, improved purification, and membrane binding. J Biol Chem 263:7136–7140

    Google Scholar 

  • Seufert W, Messer W (1987) Start sites for bidirectional in vitro replication inside the replication origin, oriC. of Escherichia coli. EMBO J 6:2469–2472

    Google Scholar 

  • Seufert W, Dobrinski B, Lurz R, Messer W (1988) Functionality of the DnaA protein binding site in DNA replication is orientation dependent. J Biol Chem 263:2719–2723

    Google Scholar 

  • Stuitje AR, de Wind N, van der Spek JC, Pors TH, Meijer M (1986) Dissection of promoter sequences involved in transcriptional activation of the Escherichia coli replication origin. Nucleic Acids Res 14:2333–2344

    Google Scholar 

  • van den Berg EA, Geerse RH, Memelink J, Bovenberg RAL, Magnée FA, van de Putte P (1985) Analysis of regulatory sequences upstream of the E. coli uvrB gene; involvement of the DnaA protein. Nucleic Acids Res 13:1829–1840

    Google Scholar 

  • Wang Q, Kaguni JM (1987) Transcriptional repression of the dnaA gene of Escherichia coli by dnaA protein. Mol Gen Genet 209:518–525

    Google Scholar 

  • Wang Q, Kaguni JM (1989) DnaA Protein regulates transcription of the rpoH gene of Escherichia coli. J Biol Chem 264:7338–7344

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by R. Devoret

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schaefer, C., Messer, W. DnaA protein/DNA interaction. Modulation of the recognition sequence. Mol Gen Genet 226, 34–40 (1991). https://doi.org/10.1007/BF00273584

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00273584

Key words

Navigation