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DNA methylation differentially enhances the expression of one of the two E. coli dnaA promoters in vivo and in vitro

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Summary

The promoter/regulatory region of the dnaA gene, whose gene product is required for the initiation of DNA replication in Escherichia coli K-12, contains an unusually large number of Dam methylation sites. In this paper we report that the expression of the dnaA gene is decreased in Dam strains of E. coli. The decrease in the expression of dnaA was measured in vivo using a dnaA-lacZ gene fusion. In vivo S1 nuclease mapping demonstrated that the decrease was due to a differential decrease in expression from the more proximal of the two dnaA promoters, dnaA2P. Comparison of the strengths of the two dnaA promoters in an in vitro transcription system using methylated and unmethylated DNA templates suggests that the effect of methylation on dnaA2P is probably at the level of RNA polymerase/DNA interaction. We suggest that this effect of methylation may be important in controlling the expression of dnaA during the E. coli cell cycle.

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References

  • Atlung T (1981) Analysis of seven dnaA suppressor loci in Escherichia coli. ICN-UCLA Symp Mol Cell Biol 21:297–314

    Google Scholar 

  • Atlung T, Rasmussen KV, Clausen E, Hansen FG (1985) Role of the dnaA protein in control of DNA replication. In: Schaechter M, Neidhardt FC, Ingraham J, Kjeldgaard NO (eds) Molecular biology of bacterial growth. Jones and Barlett Publ Inc, Portola Valley, CA: p 282–297

    Google Scholar 

  • Bagdasarian M, Izakowska M, Bagdasarian M (1977) Suppression of the DnaA phenotype by mutations in the rpoB cistron of ribonucleic acid polymerase in Salmonella typhymurium and Escherichia coli. J Bacteriol 130:577–582

    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 

  • Churchward G, Holmans P, Bremer H (1983) Increased expression of the dnaA gene has no effect on DNA replication in a dnaA + strain of Escherichia coli. Mol Gen Genet 192:506–508

    Google Scholar 

  • Eshima N, Fujii S, Honiuchi T (1972) Isolation of λind mutants. J Genet 47:125–130

    Google Scholar 

  • Fralick J (1978) Studies on the regulation of initiation of chromosome replication in Escherichia coli. J Mol Biol 122:271–286

    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 

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

    Google Scholar 

  • Hansen FG, Hansen EG, 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. The EMBO J 1:1043–1048

    Google Scholar 

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

    Google Scholar 

  • Hughes P, Squali-Houssaini FZ, Forterre P, Kohiyama M (1984) In vitro replication of a dam methylated and non-methylated oriC plasmid. J Mol Biol 176:155–159

    Google Scholar 

  • Jones PA (1985) Altering gene expression with 5-azacytidine. Cell 40:485–486

    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 

  • Kleckner N, Morisato D, Roberts D, Bender J (1984) Mechanism and regulation of Tn10 transposition. Cold Spring Harbor Symp Quant Biol 49:235–244

    Google Scholar 

  • LeGrice SFJ, Matzura H (1980) Localization of the transcription initiation site of chloramphenicol resistance gene on plasmid pAC184. FEBS Lett 113:42–46

    Google Scholar 

  • Lycett G, Orr E, Pritchard R (1980) Chloramphenicol releases a block in initiation of chromosome replication in a dnaA strain of Escherichia coli K-12. Mol Gen Genet 178:329–336

    Google Scholar 

  • Marinus MG, Morris NR (1973) Isolation of DNA methylase mutants from E. coli K-12. J Bacteriol 114:1143–1150

    Google Scholar 

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

    Google Scholar 

  • Marinus MG, Morris NR (1975) Pleiotropic effects of a DNA adenine methylation mutation (dam-3) in Escherichia coli K-12. Mutat Res 28:15–26

    Google Scholar 

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

    Google Scholar 

  • Meijer M, Messer W (1980) Functional analysis of minichromosome replication: bidirectional and unidirectional replication from the Escherichia coli replication origin, oriC. J Bacteriol 143:1049–1053

    Google Scholar 

  • Meijer M, Beck E, Hansen FG, Bergmans HEN, Messer W, von Meyenburg K, Schaller H (1979) Nucleotide sequence of the origin of replication of the Escherichia coli K-12 chromosome. Proc Natl Acad Sci USA 76:580–584

    Google Scholar 

  • Messer W, Bellekes V, Lother H (1985) Effect of dam methylation on the activity of the E. coli replication origin, oriC. The EMBO J 4:1327–1332

    Google Scholar 

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

    Google Scholar 

  • Mount DW, Low KB, Edmiston S (1972) Dominant mutations (lex) in E. coli K-12 which affect radiation sensitivity and frequency of ultraviolet light-induce mutations. J Bacteriol 112:886–893

    Google Scholar 

  • Plasterk RHA, Vollering M, Brinkman A, Van de Putte P (1984) Analysis of the methylation-regulated Mu mom transcript. Cell 36:189–196

    Google Scholar 

  • Roberts JW, Roberts CW, Craig NL (1978) E. coli recA gene product inactivates phage lambda repressor. Proc Natl Acad Sci USA 75:4714–4718

    Google Scholar 

  • Sakakibara Y, Mizukami T (1980) A temperature-sensitive Escherichia coli mutant defective inDNA replication: dnaN, a new gene adjacent to the dnaA gene. Mol Gen Genet 178:541–543

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Smith DW, Garland AM, Herman G, Enns RE, Baker TA, Zyskind JW (1985) Importance of state of methylation of oriC GATC sites in initiation of DNA replication in E. coli. The EMBO J 4:1319–1326

    Google Scholar 

  • Stuber D, Bujard H (1981) Organization of transcriptional signals in plasmids pRB322 and pACYC184. Proc Natl Acad Sci USA 78:167–171

    Google Scholar 

  • Sugimoto K, Oka A, Sugisaki H, Takanami M, Nishimura A, Yasuda Y, Hirota Y (1979) Nucleotide sequence of Escherichia coli K-12 replication origin. Proc Natl Acad Sci USA 76:575–579

    Google Scholar 

  • Szyf M, Avraham-Hactzni K, Reifman A, Shlomai J, Kaplan F, Oppenheim A, Razin A (1984) DNA methylation pattern is determined by the intracellular level of the methylase. Proc Natl Acad Sci USA 81:3278–3282

    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 

  • Wagner R, Jr, Meselson M (1976) Repair tracts in mismatched DNA heteroduplexes. Proc Natl Acad Sci USA 73:4135–4139

    Google Scholar 

  • Zyskind J, Deen L, Smith D (1977) Temporal sequence of events during the initiation process in Escherichia coli deoxyribonucleic acid replication: roles of the dnaA and dnaC gene products and ribonucleic acid polymerase. J Bacteriol 129:1466–1475

    Google Scholar 

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Communicated by N.D.F. Grindley

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Braun, R.E., Wright, A. DNA methylation differentially enhances the expression of one of the two E. coli dnaA promoters in vivo and in vitro. Mol Gen Genet 202, 246–250 (1986). https://doi.org/10.1007/BF00331644

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