Skip to main content
Log in

Autoregulation of the dnaA gene of Escherichia coli K12

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

Summary

Regulation of the dnaA gene, which codes for an essential factor for the initiation of replication from the chromosomal origin, was studied in vivo using transcriptional and translational gene fusions. We found that the dnaA gene was autoregulated over a 30-fold range by the activity of dnaA protein. Expression from the dnaA promoter region of a dnaA″lacZ fusion was inhibited up to sevenfold by surplus dnaA protein and was stimulated up to fivefold upon thermoinactivation of the mutant protein in five different dnaA(Ts) strains. The autoregulation was found to be exerted at transcription from the major dnaA promoter and was eliminated by deletion of sequences around position-65 of this promoter where a 9-bp sequence, which is also found four times in the chromosomal origin, is located.

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

  • Atlung T (1981) Analysis of seven dnaA suppressor loci in Escherichia coli. ICN-UCLA Symp Mol Cell Biol 22:297–314

    Google Scholar 

  • Atlung T (1984) Allele-specific suppression of dnaA(Ts) mutations by rpoB mutations in Escherichia coli. Mol Gen Genet 197:125–128

    Google Scholar 

  • Atlung T, Hansen FG (1983) Effect of dnaA and rpoB mutations on attenuation in the trp operon of Escherichia coli. J Bacteriol 156:985–992

    Google Scholar 

  • Atlung T, Clausen ES, Hansen FG (1984) Autorepression of the dnaA gene of Escherichia coli. In: Hübscher U (ed) Proteins involved in DNA replication. Plenum Press, New York, pp 199–208

    Google Scholar 

  • Atlung T, Rasmussen KV, Clausen ES, Hansen FG (1985) Role of the dnaA protein in control of replication. In: Schaechter M, Neidhardt FC, Ingraham J, Kjeldgaard NO (eds) Molecular biology of bacterial growth. Jones and Bartlett, Boston, in press

    Google Scholar 

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

    Google Scholar 

  • Bolivar F, Rodrigues RL, Green PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113

    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 

  • Burgers PMJ, Kornberg A, Sakakibara Y (1981) The dnaN gene codes for the β subunit of DNA polymerase III holoenzyme of Escherichia coli. Proc Natl Acad Sci USA 78:5391–5395

    Google Scholar 

  • Casadaban MJ, Cohen SN (1980) Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138:179–207

    Google Scholar 

  • Chakraborty T, Yoshinaga K, Lother H, Messer W (1982) Purification of the E. coli dnaA gene product. EMBO J 1:1545–1549

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

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

    Google Scholar 

  • Fuller RS, Kornberg A (1983) Purified dnaA protein in initiation of replication at the Escherichia coli chromosomal origin of replication. Proc Natl Acad Sci USA 80:5817–5821

    Google Scholar 

  • Fuller RS, Kaguni JM, Kornberg A (1981) Enzymatic replication of the origin of the Escherichia coli chromosome. Proc Natl Acad Sci USA 78:7370–7374

    Google Scholar 

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

    Google Scholar 

  • Gussin NG, Johnson AD, Pabo CO, Sauer RT (1983) Repressor and cro protein. Structure, function and role in lysogenization. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II. Cold Spring Harbor Laboratory, New York, pp 93–121

    Google Scholar 

  • Hansen FG, Rasmussen KV (1977) Regulation of the dnaA product in Escherichia coli. Mol Gen Genet 155:219–225

    Google Scholar 

  • Hansen FG, von Meyenburg K (1979) Characterization of the dnaA, gyrB and other genes in the dnaA region of the Escherichia coli chromosome on specialized transducing phages λtna. Mol Gen Genet 175:135–144

    Google Scholar 

  • Hansen FG, Koefoed S, von Meyenburg K, Atlung T (1981) Transcription and translation events in the oriC region of the E. coli chromosome. ICN-UCLA Symp Mol Cell Biol 22:37–55

    Google Scholar 

  • Hansen EB, Hansen FG, von Meyenburg K (1982) The nucleotide sequence of the dnaA gene and the first part of the dnaA gene of Escherichia coli K-12. Nucleic Acids Res 10:7373–7385

    Google Scholar 

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

    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 Escherichia coli. Mol Gen Genet 156:387–396

    Google Scholar 

  • Hirota Y, Mordoh J, Jacob F (1970) On the process of cellular division in Escherichia coli III. Thermosensitive mutants of Escherichia coli altered in the process of DNA initiation. J Mol Biol 53:369–387

    Google Scholar 

  • Kellenberger-Gujer G, Podhajska AJ, Caro L (1978) A cold sensitive dnaA mutant of E. coli which overinitiates chromosome replication at low temperature. Mol Gen Genet 162:9–16

    Google Scholar 

  • Kimura M, Yura T, Nagata T (1980) Isolation and characterination of Escherichia coli dnaA amber mutants. J Bacteriol 144:649–655

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lark KG (1972) Evidence for direct involvement of RNA in the initiation of DNA replication in E. coli 15T-. J Mol Biol 64:47–60

    Google Scholar 

  • Lother H, Bukh H-J, Morelli G, Heimann B, Chakraborty T, Messer W (1981) Genes, transcriptional units and functional sites in and around the E. coli replication origin. ICN-UCLA Symp Mol Cell Biol 22:57–77

    Google Scholar 

  • Mandel M, Higa A (1970) Calcium dependent bacteriophage DNA injection. J Mol Biol 53:159–162

    Google Scholar 

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

    Google Scholar 

  • Marinus MG, Morris NR (1974) Biological function for the 6-methyladenine residues in the DNA of Escherichia coli K-12. J Mol Biol 85:309–322

    Google Scholar 

  • Marsh RC, Worcel A (1977) A DNA fragment containing the origin of replication of the Escherichia coli chromosome. Proc Natl Acad Sci USA 74:2720–2724

    Google Scholar 

  • Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560

    Google Scholar 

  • Meyenburg K von, Hansen FG, Riise E, Bergmans HEN, Meijer M, Messer W (1979) Origin of replication, oriC, of the Escherichia coli K12 chromosome: Genetic mapping and minichromosome replication. Cold Spring Harbor Symp Quant Biol 43:121–128

    Google Scholar 

  • Meyenburg K von, Jørgensen BB, Nielsen J, Hansen FG (1982) Promoters of the atp operon coding for the membrane bound ATP synthase of Escherichia coli mapped by Tn10 insertion mutations. Mol Gen Genet 188:240–248

    Google Scholar 

  • Meyenburg K von, Hansen FG, Atlung T, Boe L, Clausen IG, van Deurs B, Hansen EB, Jørgensen BB, Jørgensen F, Koppes L, Michelsen O, Nielsen J, Pedersen PE, Rasmussen KV, Riise E, Skovgaard O (1985) Facets of the chromosomal origin of replication, oriC, of Escherichia coli. In: Schaechter M, Neidhardt FC, Ingraham J, Kjeldgaard NO (eds) Molecular biology of bacterial growth. Jones and Bartlett, Boston, in press

    Google Scholar 

  • Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Neidhardt FC, Bloch PL, Pedersen S, Reeh SV (1977) Chemical measurement of steady state levels of ten aminoacyl-transfer ribonucleic acid synthetases in Escherichia coli. J Bacteriol 129:378–387

    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 

  • Pedersen S, Reeh SV, Watson RJ, Friesen JD, Fiil NP (1976) Analysis of the proteins synthesized in ultraviolet light-irradiated Escherichia coli following infection with the bacteriophages λdrifD18 and λdfus-3. Mol Gen Genet 144:339–343

    Google Scholar 

  • Polaczek P, Ciesla Z (1984) Effect of altered efficiency of the RNA I and RNA II promoters on in vivo replication of ColE1-like plasmids in Escherichia coli. Mol Gen Genet 194:227–231

    Google Scholar 

  • Remaut E, Stanssens P, Fiers W (1981) Plammid vectors for high efficiency expression by the pL promotor of coliphage lambda. Gene 15:81–93

    Google Scholar 

  • Sakakibara Y, Tsukano H, Sako T (1981) Organization and transcription of the dnaA and dnaN genes of Escherichia coli. Gene 13:47–55

    Google Scholar 

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

    Google Scholar 

  • Schaus N, O'Day K, Peters W, Wright A (1981a) Isolation and characterization of amber mutations in gene dnaA of Escherichia coli K-12. J Bacteriol 145:904–913

    Google Scholar 

  • Schaus N, O'Day K, Wright A (1981b) Suppression of amber mutations in the dnaA gene of Escherichia coli K-12 by secondary mutations in rpoB. ICN-UCLA Symp Mol Cell Biol 22:315–323

    Google Scholar 

  • Schultz J, Silhavy TJ, Berman ML, Fiil NP, Emr SD (1982) A previously unidentified gene in the spc operon of Escherichia coli K12 specifies a component of the protein export machinery. Cell 31:227–235

    Google Scholar 

  • Siebenlist U, Simpson RB, Gilbert W (1980) Escherichia coli RNA polymerase interacts homologously with two different promoters. Cell 20:269–281

    Google Scholar 

  • Stoker NG, Fairweather NF, Spratt BG (1982) Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene 18:335–341

    Google Scholar 

  • Stüber D, Bujard H (1981) Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci USA 78:167–171

    Google Scholar 

  • Stüber D, Bujard H (1982) Transcription from efficient promoters can interfere with plasmid replication and diminish expression of plasmid specified genes. EMBO J 1:1399–1404

    Google Scholar 

  • Sutcliffe JG (1979) Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43:77–90

    Google Scholar 

  • Tinocho I, Borer PN, Dengler B, Levine MD, Uhlenbeck OC, Crothers DM, Gralla J (1973) Improved estimation of secondary structure in ribonucleic acids. Nature (New Biology) 246:40–41

    Google Scholar 

  • Tippe-Schindler R, Zahn G, Messer W (1979) Control of the initiation of DNA replication in Escherichia coli. I. Negative control of initiation. Mol Gen Genet 168:185–195

    Google Scholar 

  • Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268

    Google Scholar 

  • Zyskind JW, Harding NE, Takeda Y, Cleary JM, Smith DW (1981) The DNA replication origin of the Enterobacteriaceae. ICN-UCLA Symp Mol Cell Biol 22:13–25

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by H. Saedler

Rights and permissions

Reprints and permissions

About this article

Cite this article

Atlung, T., Clausen, E.S. & Hansen, F.G. Autoregulation of the dnaA gene of Escherichia coli K12. Molec. Gen. Genet. 200, 442–450 (1985). https://doi.org/10.1007/BF00425729

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00425729

Keywords

Navigation