Skip to main content
SpringerLink
Log in

Expression and regulation of a dnaA homologue isolated from Pseudomonas putida

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

Summary

A gene homologous to the Escherichia coli dnaA gene was isolated from Pseudomonas putida and its transcription was investigated in E. coli as well as in P. putida. In both species the P. putida dnaA gene is transcribed from two promoters, one of which shows strong homology to promoters recognized by the α54 factor found in both bacteria. In E. coli transcription of the P. putida dnaA gene can be repressed by overproduction of E. coli DnaA protein, presumably due to the presence of several DnaA-box-like sequences found in the promoter region. Likewise the P. putida DnaA protein is able to regulate expression of the E. coli dnaA gene but we failed to demonstrate autoregulation of the P. putida dnaA gene. A point mutation was introduced into the P. putida dnaA gene, equivalent to the ATP binding site mutation present in E. coli dnaA5 and dnaA46 mutants, and this alteration abolished the ability of the protein to repress the expression of the E. coli dnaA gene. These results indicate that DnaA proteins from other species than E. coli have maintained the ability to recognize the DnaA box sequence and that the conservation between the DnaA proteins reflects functionally similar domains.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Andrup L, Atlung T, Ogasawara N, Yoshikawa H, Hansen FG (1988) Interaction of the Bacillus subtilis DnaA-like protein with Escherichia coli DnaA protein. J Bacteriol 170:1333–1338

    Google Scholar 

  • Atlung T, Clausen ES, Hansen FG (1985) Autoregulation of the dnaA gene of Escherichia coli K12. Mol Gen Genet 200:442–450

    Google Scholar 

  • Bachmann BJ (1987) Linkage map of Escherichia coli K-12. In: Neidhardt FC, Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium: Cellular and Molecular biology. American Society for Microbiology, Washington DC, pp 807–876

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Braun RE, O'Day K, Wright A (1985) Autoregulation of the DNA replication gene dnaA in E. coli K-12. Cell 40:159–169

    Google Scholar 

  • Casadaban MJ, Cohen SN (1980) Analysis of gene control signals by DNA fusion and cloning in E. coli. J Mol Biol 138:179–208

    Google Scholar 

  • Clark DJ, Maaløe O (1967) DNA replication and the division cycle in Escherichia coli. J Mol Biol 23: 99–112

    Google Scholar 

  • Dairi T, Inokuchi K, Mizuno T, Mizushima S (1985) Positive control of transcription initiation in Escherichia coli: A base substitution at the Pribnow box renders ompF expression independent of a positive regulator. J Mol Biol 184:1–6

    Google Scholar 

  • deBoer HA, Comstock LJ, Vasser M (1983) The tac promoter: A functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci USA 80:21–25

    Google Scholar 

  • Dixon R (1986) The xylABC promoter from the Pseudomonas putida TOL plasmid is activated by nitrogen regulatory genes in E. coli. Mol Gen Genet 203:129–136

    Google Scholar 

  • Fujita MQ, Yoshikawa H, Ogasawara N (1989) Structure of the dnaA region of Pseudomonas putida: Conservation among three bacteria, Bacillus subtilis, Escherichia coli and Pseudomonas putida. Mol Gen Genet 215:381–387

    Google Scholar 

  • Fujita MQ, Yoshikawa H, Ogaswara N (1990) Structure of the dnaA region of Micrococcus luteus: conservation and variation among eubacteria. Gene 93:73–78

    Google Scholar 

  • Fürste JP, Pansegrau W, Frank R, Blocker H, Stolz P, Bagdasarian M, Lanka E (1986) Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene 48:119–131

    Google Scholar 

  • Gragerov AJ, Chenchik AA, Aivasashvilli VA, Beabealashvilli RS, Nikiforov VG (1984) Escherichia coli and Pseudomonas putida RNA polymerases display identical contacts with promoters. Proc Natl Acad Sci USA 195:511–515

    Google Scholar 

  • Hansen EB, Atlung T, Hansen FG, Skovgaard O, von Meyenburg K (1984) Fine structure genetic map and complementation analysis of mutations in the dnaA gene of Escherichia coli. Mol Gen Genet 196:387–396

    Google Scholar 

  • Hansen FG, Hansen EB, Atlung T (1982) The nucleotide sequence of the dnaA gene promoter and of the adjacent rpmH gene, coding for the ribosomal protein L34, of Escherichia coli. EMBO J 1:1043–1048

    Google Scholar 

  • Hansen FG, Koefoed S, Sørensen L, Atlung T (1987) Titration of the DnaA protein by oriC dnaA boxes increases dnaA gene expression in Escherichia coli. EMBO J 6:255–258

    Google Scholar 

  • Hansen FG, Atlung T, Braun RE, Wright A, Hughes P, Kohiyama M (1991) Initiator (DnaA) protein concentration as a function of growth rate in Escherichia coli and Salmonella typhimurium. J Bacteriol 173:5194–5199

    Google Scholar 

  • Inouye S, Asai Y, Nakazawa A, Nakazawa T (1986) Nucleotide sequence of a DNA fragment promoting transcription in Pseudomonas putida. J Bacteriol 166:739–745

    Google Scholar 

  • Jeenes DJ, Soldati JL, Baur H, Watson JM, Mercenier A, Reimmann C, Lesinger T, Haas D (1986) Expression of biosynthetic genes from Pseudomonas aeruginosa and Escherichia coli in the heterologous host. Mol Gen Genet 203:421–429

    Google Scholar 

  • Kohler T, Harayama S, Ramos J-L, Timmis KN (1989) Involvement of Pseudomonas putida RpoN sigma factor in regulation of various metabolic functions. J Bacteriol 171:4326–4333

    Google Scholar 

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

    Google Scholar 

  • Kunkel TA (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492

    Google Scholar 

  • Lindahl G, Lindahl T (1984) Initiation of RNA replication in Escherichia coli: RNascH deficient mutants do not require the dnaA protein. Mol Gen Genet 196:283–289

    Google Scholar 

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

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • McCleary JA, Witney F, Geisselsoder J (1989) Efficient site-directed in vitro mutagenesis using phagemid vectors. BioTechniques 7:282–289

    Google Scholar 

  • Mead DA, Szczesna-Skorupa E, Kemper B (1986) Single-stranded DNA ‘blue’ T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng 1:67–74

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Moriya S, Ogasawara N, Yoshikawa H (1985) Structure and function of the region of the replication origin of the Bacillus subtilis chromosome, III Nucleotide sequence of some 10,000 base pairs in the origin region. Nucleic Acids Res 13:2251–2265

    Google Scholar 

  • Moriya S, Kato K, Yoshikawa H, Ogasawara N (1990) Isolation of a dnaA mutant of Bacillus subtilis defective in initiation of replication: Amount of dnaA protein determines cells initiation potential. EMBO J 9:2905–2910

    Google Scholar 

  • Ohmori H, Kimura M, Nagata T, Sakakibara Y (1984) Structural analysis of the dnaA and dnaN genes of Escherichia coli. Gene 28:159–170

    Google Scholar 

  • Quinones A, Messer W (1988) Discoordinate gene expression in the dnaA-dnaN operon of Escherichia coli. Mol Gen Genet 213:118–124

    Google Scholar 

  • Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353

    Google Scholar 

  • Sakakibara Y, Yuasa S (1982) Continuous synthesis of the dnaA gene product of Escherichia coli in the cell cycle. Mol Gen Genet 186:87–94

    Google Scholar 

  • Sako T, Sakabibara Y (1980) Coordinate expression of the Escherichia coli dnaA and dnaN genes. Mol Gen Genet 179:521–526

    Google Scholar 

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

    Google Scholar 

  • Sancar A, Hack AM, Rupp WD (1979) Simple method for identification of plasmid coded proteins. J Bacteriol 137:692–693

    Google Scholar 

  • Schneider K, Beck CF (1987) New expression vectors for identifying and testing signal structures for initiation and termination of transcription. Methods Enzymol 153:452–461

    Google Scholar 

  • Sinden RR, Carlson JO, Pettijohn DE (1980) Torsional tension in the DNA double helix measured with trimethylpsoralen in living E. coli cells: analogous measurements in insect and human cells. Cell 21:773–783

    Google Scholar 

  • Skovgaard O (1990) Nucleotide sequence of a Proteus mirabilis fragment homologous to the 60K-rnpA-dnaA-dnaN-recF-gyrB region of Escherichia coli. Gene 93:27–34

    Google Scholar 

  • Skovgaard O, Hansen FG (1987) Comparison of dnaA nucleotide sequences of Escherichia coli, Salmonella typhimurium, and Serratia marcescens. J Bacteriol 169:3976–3981

    Google Scholar 

  • Vieira J, Messing J (1987) Production of single-stranded plasmid DNA. Methods Enzymol 153:3–11

    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) A novel sigma factor is involved in expression of the rpoH gene of Escherichia coli. J Bacteriol 171:4248–4253

    Google Scholar 

  • Yee T, Smith DW (1990) Pseudomonas chromosomal replication origins: A bacterial class distinct from Escherichia coli-type origins. Proc Natl Acad Sci USA 87:1278–1282

    Google Scholar 

Download references

Author information

Author notes

  1. Hanne Ingmer

    Present address: Department of Genetics, Stanford University School of Medicine, 94305, Stanford, CA, USA

Authors and Affiliations

  1. Department of Microbiology, The Technical University of Denmark, Building 221, DK-2800, Lyngby, Denmark

    Hanne Ingmer & Tove Atlung

Authors
  1. Hanne Ingmer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tove Atlung
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by R. Devoret

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ingmer, H., Atlung, T. Expression and regulation of a dnaA homologue isolated from Pseudomonas putida . Molec. Gen. Genet. 232, 431–439 (1992). https://doi.org/10.1007/BF00266248

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00266248

Key words

Search

Navigation