Skip to main content
Log in

Recombination between repeats in Escherichia coli by a recA-independent, proximity-sensitive mechanism

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

Abstract

We have examined the influence of proximity on the efficiency of recombination between repeated DNA sequences in Escherichia coli. Our experiments have employed a plasmid-based assay to detect deletions between direct repeats of 100 bp. The rate of deletion of the juxtaposed direct repeats was reasonably high at 6 × 10−5 per cell. A comparison of recA+ and recA mutant strains showed that these deletion events are primarily the result of recA-independent recombination at these homologous sequences. Random restriction fragments of yeast or E. coli genomic DNA were used to separate the two repeats. Deletion rates decreased over two orders of magnitude with increasing separation of up to 7 kb. There was a surprisingly strong effect of even short sequence separations, with insertions of a few hundred base pairs exhibiting 10-fold reductions of deletion rates. No effect of recA on the efficiency of deletion was observed at any distance between repeats.

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

  • Albertini AM, Hofer M, Calos MP, Miller JH (1982a) On the formation of spontaneous deletion: the importance of short sequence homologies in the generation of large deletions. Cell 29:319–328

    Google Scholar 

  • Albertini AM, Hofer M, Calos MP, Tlsty TD, Miller JH (1982b) Analysis of spontaneous deletions and gene amplification in the lac region of Escherichia coli. Cold Spring Harbor Symp Quant Biol 47:841–850

    Google Scholar 

  • Allgood ND, Silhavy TJ (1988) Illegitimate recombination in bacteria. In: Kucherlapati R, Smith GR (eds) Genetic recombination. American Society for Microbiology, Washington, DC, pp 309–330

    Google Scholar 

  • Ausubel FA, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1989). Short protocols in molecular biology. Wiley, New York

    Google Scholar 

  • Bachmann BJ (1987) Derivations and genotypes of some mutant derivatives of Escherichia coli K-12. In: Neidhardt FD (ed) Escherichia coli and Salmonella typhimurium: cellular and molecular biology. American Society for Microbiology, Washington, DC, pp 1190–1219

    Google Scholar 

  • Bi X, Liu LF (1994) recA-independent and recA-dependent intramolecular plasmid recombination: differential homology requirement and distance effect. J Mol Biol 235:414–423

    Google Scholar 

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

    Google Scholar 

  • Chung CT, Niemela SL, Miller RH (1989) One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci USA 86:2172–2175

    Google Scholar 

  • Csonka L, Clark AJ (1979) Deletions generated by the transposon Tn10 in the srl recA region of the Escherichia coli K-12 chromosome. Genetics 93:321–343

    Google Scholar 

  • Dasgupta U, Weston-Hafer K, Berg DE (1987) Local DNA sequence control of deletion formation in Escherichia coli plasmid pBR322. Genetics 115:41–49

    Google Scholar 

  • Dianov GL, Kuzminov AV, Mazin AV, Salganik RI (1991) Molecular mechanisms of deletion formation in Escherichia coli plasmids I. Deletion formation mediated by long direct repeats. Mol Gen Genet 228:153–159

    Google Scholar 

  • Dower WJ, Miller JF, Ragsdale CW (1988) High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16:6127–6145

    Google Scholar 

  • Dugaiczyk A, Boyer HW, Goodman HM (1975) Ligation of EcoRI endonuclease-generated DNA fragments into linear and circular structures. J Mol Biol 96:171–184

    Google Scholar 

  • Efstratiadis A, Psalony JW, Maniatis T, Lawn RM, O'Connell C, Spritz RA, DeRiel JK, Forget BG, Weissman SM, Slightom JL, Blechl AE, Smithies O, Baralle FE, Shoulders CC, Proudfoot NJ (1980) The structure and evolution of the human β-globin gene family. Cell 21:653–668

    Google Scholar 

  • Ehrlich SD (1989) Illegitimate recombination in bacteria. In: Berg DE, Howe MM (eds) Mobile DNA. American Society for Microbiology, Washington, DC, pp 799–832

    Google Scholar 

  • Farabaugh PJ, Schmeissner U, Hofer M, Miller JH (1978) Genetic studies of the lac repressor. VII. On the molecular nature of spontaneous hotspots in the lacI gene of Escherichia coli. J Mol Biol 126:847–857

    Google Scholar 

  • Fasman GD (1975). Handbook of biochemistry and molecular biology, 3rd edn. CRC Press, Cleveland, Ohio

    Google Scholar 

  • Galas DJ (1978) An analysis of sequence repeats in the lacI gene of Escherichia coli. J Mol Biol 126:858–863

    Google Scholar 

  • Gillen J, Willis DK, Clark AJ (1981) Genetic analysis of the RecE pathway of genetic recombination in Escherichia coli K-12. J Bacteriol 145:521–532

    Google Scholar 

  • Glickman BW, Ripley S (1984) Structural intermediates of deletion mutagenesis: a role for palindromic DNA. Proc Natl Acad Sci USA 81:512–516

    Google Scholar 

  • King SR, Richardson JP (1986) Role of homology and pathway specificity for recombination between plasmids and bacteriophage λ. Mol Gen Genet 204:141–147

    Google Scholar 

  • Lea DE, Coulson CA (1949) The distribution of the numbers of mutants in bacterial populations. J Genetics 49:264–285

    Google Scholar 

  • Lovett ST, Drapkin PT, Sutera VA Jr., Gluckman-Peskind TJ (1993) A sister-strand exchange mechanism for recA-independent deletion of repeated DNA sequences in Escherichia coli. Genetics 135:631–642

    Google Scholar 

  • Mahajan SK (1988) Pathways of homologous recombination in Escherichia coli. In: Kucherlapati R, Smith GR (eds) Genetic recombination. American Society for Microbiology, Washington, DC, pp 87–140

    Google Scholar 

  • Matfield M, Badawi R, Brammar WJ (1985) Rec-dependent and Rec-independent recombination of plasmid-borne duplications in Escherichia coli K12. Mol Gen Genet 199:518–523

    Google Scholar 

  • Mazin AV, Kuzminov AV, Dianov GL, Salganik RI (1991) Molecular mechanisms of deletion formation in Escherichia coli plasmids II. Deletion formation mediated by short direct repeats. Mol Gen Genet 228:209–214

    Google Scholar 

  • Meuth M (1989) Illegitimate recombination in mammalian cells. In: Berg DE, Howe MM (eds) Mobile DNA. American Society for Microbiology, Washington, DC, pp 833–860

    Google Scholar 

  • Shen P, Huang HV (1986) Homologous recombination in Escherichia coli: dependence on substrate length and homology. Genetics 112:441–457

    Google Scholar 

  • Sherman F, Fink GR, Hicks JB (1983). Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Sinden RR, Zheng G, Brankamp RG, Allen KN (1991) On deletion of inverted repeated DNA in Escherichia coli: effects of length, thermal stability, and cruciform formation in vivo. Genetics 129:991–1005

    Google Scholar 

  • Singer BS, Westlye J (1988) Deletion formation in bacteriophage T4. J Mol Biol 202:233–243

    Google Scholar 

  • Streisinger G, Okada Y, Emrich J, Newton J, Tsugita A, Terzaghi E, Inouye I (1966) Frameshift mutations and the genetic code. Cold Spring Harbor Symp Quant Biol 31:77–86

    Google Scholar 

  • Walker GC (1987) The SOS response of Escherichia coli. In: Neidhardt FD (ed) Escherichia coli and Salmonella typhimurium: cellular and molecular biology. American Society for Microbiology, Washington, DC, pp 1346–1357

    Google Scholar 

  • Watt VM, Ingles CJ, Urdea MS, Rutter WJ (1985) Homology requirements for recombination in Escherichia coli. Proc Natl Acad Sci USA 82:4768–4772

    Google Scholar 

  • Weston-Hafer D, Berg DE (1989) Palindromy and the location of deletion endpoints in Escherichia coli. Genetics 121:651–658

    Google Scholar 

  • Weston-Hafer K, Berg DE (1991) Deletions in plasmid pBR322: replication slippage involving leading and lagging strands. Genetics 127:649–6555

    Google Scholar 

  • Willetts NS, Clark AJ, Low B (1969) Genetic location of certain mutations conferring recombination deficiency in Escherichia coli. J Bacteriol 97:244–249

    Google Scholar 

  • Yi T-M, Stearns D, Demple B (1988) Illegitimate recombination in an Escherichia coli plasmid: modulation by DNA damage and a new bacterial gene. J Bacteriol 170:2898–2903

    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

Lovett, S.T., Gluckman, T.J., Simon, P.J. et al. Recombination between repeats in Escherichia coli by a recA-independent, proximity-sensitive mechanism. Molec. Gen. Genet. 245, 294–300 (1994). https://doi.org/10.1007/BF00290109

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00290109

Keywords

Navigation