Skip to main content
SpringerLink
Log in

Expression of thednaB gene ofEscherichia coli is inducible by replication-blocking DNA damage in arecA-independent manner Received: 4 April 1995

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

Abstract

The replicative DNA helicase encoded by thednaB gene is essential for chromosomal DNA replication inEscherichia coli. The DnaB protein is a component of the σX-type primosome which is regarded as a model system for lagging strand synthesis of the chromosome. Using translationallacZ fusions at the plasmid and chromosomal levels, we studied the influence of DNA-damaging agents ondnaB gene expression. We found that DNA damage caused by mitomycin C, methyl methanesulphonate, 4-nitroquinoline N-oxide, and UV irradiation led to a moderate, but significant induction ofdnaB gene expression. Comparative S1 analysis of transcripts in untreated and induced cells demonstrated that the induction is due to increased transcription from thednaB promoter. In contrast to other DNA damage-inducible replication genes, such asdnaA,dnaN,dnaQ, andpolA, expression of which is not inducible inrecA andlexA mutants, the induction ofdnaB was also observed in arecA1 mutant. These results show that the induction ofdnaB gene expression by replication-blocking DNA damage is due to a mechanism other than the indirectly SOS-dependent induction of the other DNA replication genes. Moreover, the data suggest that replication proteins are involved in recovery from replication-blocking DNA damage in two different ways — on the one hand at the level of initiation and on the other hand at the level of elongation.

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

  • Allen GC, Kornberg A (1991) Fine balance in the regulation of DnaB helicase by DnaC protein in replication inEscherichia coli. J Biol Chem 266:22096–22101

    Google Scholar 

  • Allen GC, Kornberg A (1993) Assembly of the primosome of DNA replication inEscherichia coli. J Biol Chem 268:19204–19209

    Google Scholar 

  • Allen GC, Dixon NE, Kornberg A (1993) Strand switching of a replicative DNA helicase promoted by theE. coli primosome. Cell 74:713–722

    Google Scholar 

  • Arai K, Kornberg A (1979) A general priming system employing only DnaB protein and primase for DNA replication. Proc Natl Acad Sci USA 76:4308–4312

    Google Scholar 

  • Arai K, Kornberg A (1981a) Mechanism ofdnaB protein action. II. ATP hydrolysis bydnaB protein dependent on single- or doublestranded DNA. J Biol Chem 256:5253–5259

    Google Scholar 

  • Arai K, Kornberg A (1981b) Mechanism ofdnaB protein action. III. Allosteric role of ATP in the alteration of structure by DnaB protein in priming replication. J Biol Chem 256:5260–5266

    Google Scholar 

  • Arai K, Kornberg A (1981c) Mechanism ofdnaB protein action. IV. General priming of DNA replication bydnaB protein and primase compared with RNA polymerase. J Biol Chem 256: 5267–5272

    Google Scholar 

  • Arai K, Low R, Kobori J, Shlomai J, Kornberg A (1981) Mechanism ofdnaB protein interaction. V. Association ofdnaB protein, protein n', and other prepriming proteins in the primosome of DNA replication. J Biol Chem 256:5273–5280

    Google Scholar 

  • Bachmann B (1990) Linkage map ofEscherichia coli. Edition 8. Microbiol Rev 54:130–197

    Google Scholar 

  • Bagg A, Kenyon CJ, Walker GC (1981) Inducibility of a gene product required for UV and chemical mutagenesis inEscherichia coli. Proc Natl Acad Sci USA 78:5749–5753

    Google Scholar 

  • Baker T, Sekimizu K, Funnell B, Kornberg A (1986) Extensive unwinding of the plasmid template during staged enzymatic initiation of DNA replication from the origin of theEscherichia coli chromosome. Cell 45:53–64

    Google Scholar 

  • Baker T, Funnel B, Kornberg A (1987) Helicase action ofdnaB protein during replication from theE. coli chromosomal origin in vitro. J Biol Chem 262: 6877–6885

    Google Scholar 

  • Basu AK, Hanrahan CJ, Malia SA, Kumart S, Bizanek R, Tomasz M (1993) Effect of site-specifically located mitomycin C-DNA monoadducts on in vitro DNA synthesis by DNA polymerases. Biochemistry 32:4708–4718

    Google Scholar 

  • Berk AJ, Sharp PA (1977) Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell 12:721–732

    Google Scholar 

  • Boiteux S, Huisman O, Laval J (1984) 3-Methyladenine residues in DNA induce the SOS functionsfiA inEscherichia coli. EMBO J 3:2569–2573

    Google Scholar 

  • Brosius J, Cate R, Perlmutter P (1982) Precise location of two promoters for theβ-lactamase gene of pBR322. J Biol Chem 257:9205–9210

    Google Scholar 

  • Diederich L, Rasmussen LJ, Messer W (1992) New cloning vectors for integration into the λ attachment siteattB of theEscherichia coli chromosome. Plasmid 28:14–24

    Google Scholar 

  • Gocke E (1991) Mechanism of quinolone mutagenicity in bacteria. Mutat Res 248:135–143

    Google Scholar 

  • Iyer VN, Szybalski W (1964) Mitomycins and porfiromycin: chemical mechanism of activation and cross-linking of DNA. Science 145:55–58

    Google Scholar 

  • Jeggo P, Defais M, Samson L, Schendel P (1977) An adaptive response of E. coli to low levels of alkylating agents: comparison with previously characterized DNA repair pathways. Mol Gen Genet 157:1–9

    Google Scholar 

  • Kaasch M, Kaasch J, Quifiones A (1989) Expression of thednaN anddnaQ genes ofEscherichia coli is inducible by mitomycin C. Mol Gen Genet 219:187–192

    Google Scholar 

  • Kaguni J, Kornberg A (1984) Replication initiated at the origin (oriC) of theE. coli chromosome reconstituted with purified enzymes. Cell 38:183–190

    Google Scholar 

  • Kobori JA, Kornberg A (1982a) TheEscherichia coli dnaC gene product. I. Overproduction of the dnaC proteins ofEscherichia coli andSalmonella typhimurium by cloning into a high copy number plasmid. J Biol Chem 257:13757–13762

    Google Scholar 

  • Kobori JA, Kornberg A (1982b) TheEscherichia coli dnaC gene product. III. Properties of the dnaB-dnaC protein complex. J Biol Chem 257:13770–13775

    Google Scholar 

  • Kornberg A, Baker TA (1992) DNA replication, 2nd edn. Freeman. New York

    Google Scholar 

  • LeBowitz J, McMacken R (1986) TheEscherichia coli dnaB replication protein is a DNA helicase. J Biol Chem 261:8778–8784

    Google Scholar 

  • Lindahl T, Sedgwick B (1988) Regulation and expression of the adaptive response to alkylating agents. Annu Rev Biochem 57:133–157

    Google Scholar 

  • Little JW, Mount DW (1982) The SOS regulatory system ofEscherichia coli. Cell 29:11–22

    Google Scholar 

  • Liu LF (1989) DNA topoisomerase poisons as antitumor drugs. Annu Rev Biochem 58:351–375

    Google Scholar 

  • Masai H, Arai K (1988) Operon structure ofdnaT anddnaC genes essential for normal and- stable replication ofEscherichia coli chromosome. J Biol Chem 263:15083–15093

    Google Scholar 

  • Masai H, Bond MW, Arai K (1986) Cloning of theEscherichia coli gene for primosomal proteini: the relationship todnaT, essential for chromosomal DNA replication. Proc Natl Acad Sci USA 83:1256–1260

    Google Scholar 

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

    Google Scholar 

  • Minton NP (1984) Improved plasmid vectors for the isolation of translational lac gene fusions. Gene 31: 269–273

    Google Scholar 

  • Nakayama N, Arai N, Bond MW, Kaziro Y, Arai K (1984) Nucleotide sequence ofdnaB and the primary structure of thednaB protein fromEscherichia coli. J Biol Chem 259:97–101

    Google Scholar 

  • Otsuka M, Nakabeppu Y, Sekiguchi M (1985) Ability of various alkylating agents to induce adaptive and SOS responses: a study withlacZ fusions. Mutat Res 146:149–154

    Google Scholar 

  • Quillardet P, Hofnung M (1985) The SOS chromotest, a colorometric bacterial assay for genotoxins: procedures. Mutat Res 147:159–170

    Google Scholar 

  • Quifiones A, Piechocki R, Messer W (1988) Expression of theEscherichia coli dnaQ (mutD) gene is inducible. Mol Gen Genet 211:106–112

    Google Scholar 

  • Quifiones A, Kaasch J, Kaasch M, Messer W (1989) Induction ofdnaN anddnaQ gene expression in Escherichia coli by alkylation damage to DNA. EMBO J 8:587–593

    Google Scholar 

  • Quifiones A, Jüterbock W-R, Messer W (1991a) Expression of thednaA gene ofEscherichia coli is inducible by DNA damage. Mol Gen Genet 227:9–16

    Google Scholar 

  • Quifiones A, Jüterbock W-R, Messer W (1991b) DNA lesions that block DNA replication are responsible for thednaA induction caused by DNA damage. Mol Gen Genet 231:81–87

    Google Scholar 

  • Sambrook J, Maniatis T, Fritsch EF (1989) Molecular cloning — a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Samson L, Cairns J (1977) A new pathway for DNA repair inEscherichia coli. Nature 267:281–283

    Google Scholar 

  • Schekman R, Weiner J, Weiner A, Kornberg A (1975) Ten proteins required for conversion of σX174 single-stranded DNA to duplex formin vitro: resolution and reconstitution. J Biol Chem 250:5859–5865

    Google Scholar 

  • Silhavy TJ, Berman ML, Enquist LW (1984) Experiments with gene fusions. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Singer B, Kusmierek T (1982) Chemical mutagenesis. Annu Rev Biochem 51:655–693

    Google Scholar 

  • Thomas DC, Husain I, Chaney SG, Panigrahi GB, Walker IG (1991) Sequence effect on incision by (A)BC excinuclease of 4NQO adducts and UV photoproducts. Nucleic Acids Res 19:365–370

    Google Scholar 

  • Tomasz M, Chawla AK, Lipman R (1988) Mechanism of monofunctional and bifunctional alkylation of DNA by mitomycin C. Biochemistry 27: 3182–3187

    Google Scholar 

  • Volkert MR (1988) Adaptive response ofscherichia coli to alkylation damage. Environ Mol Mutat 11:241–255

    Google Scholar 

  • Wahle E, Lasken RS, Kornberg A (1989) The dnaB-dnaC replication protein complex ofEscherichia coli. I. Formation and properties. J Biol Chem 264:2463–2468

    Google Scholar 

  • Walker GC (1984) Mutagenesis and inducible responses to DNA damage inEscherichia coli. Microbiol Rev 48:60–93

    Google Scholar 

  • Walker GC (1985) Inducible DNA repair systems. Annu Rev Biochem 54:425–457

    Google Scholar 

  • Walker GC (1987) The SOS response ofEscherichia coli. In: Neidhardt FC, Ingraham JL, Low KB, Magasanik B, Schaechter M, Umbarger HE (eds)Escherichia coli andSalmonella typhimurium. Cellular and molecular biology. American Microbiology Society, Washington DC, pp 1346–1357

    Google Scholar 

  • Wandt G, Quifiones A (1991) Expression of thepolA gene ofEscherichia coli is inducible by DNA damage and DnaA protein overproduction. Biol Chem Hoppe-Seyler 372:775

    Google Scholar 

  • Wickner S, Hurwitz J (1974) Conversion of σX174 viral DNA to double-stranded form by purifiedEscherichia coli proteins. Proc Natl Acad Sci USA 71:4120–4124

    Google Scholar 

  • Wickner S, Hurwitz J (1975) Interaction of Escherichia colidnaB anddnaC(D) gene products in vitro. Proc Natl Acad Sci USA 72:921–925

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Genetik, Martin-Luther-Universität Halle-Wittenberg, Domplatz l, D 06108, Halle (Saale), Germany

    Sabine Kleinsteuber & Ariel Quiñones

Authors
  1. Sabine Kleinsteuber
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ariel Quiñones
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by H. Böhme

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kleinsteuber, S., Quiñones, A. Expression of thednaB gene ofEscherichia coli is inducible by replication-blocking DNA damage in arecA-independent manner Received: 4 April 1995. Molec. Gen. Genet. 248, 695–702 (1995). https://doi.org/10.1007/BF02191709

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02191709

Key words

Search

Navigation