• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Jul 1996; 178(13): 3840–3845.
PMCID: PMC232644

Plasmid recombination by the RecBCD pathway of Escherichia coli.

Abstract

Previously, we demonstrated that exonuclease I-deficient strains of Escherichia coli accumulate high-molecular-weight linear plasmid concatemers when transformed with plasmids carrying the chi sequence (5'- GCTGGTGG-3') (M. M. Zaman and T. C. Boles, J. Bacteriol. 176:5093-5100, 1994). Since high-molecular weight linear DNA is believed to be the natural substrate for RecBCD-mediated recombination during conjugation (A. J. Clark and K. B. Low, p. 155-215, in K. B. Low, ed., The Recombination of Genetic Material, 1988), we analyzed the recombination frequencies of chi+ and chi0 plasmids in sbcB strains. Here, we report that chi sites stimulate plasmid recombination frequency by 16-fold in sbcB strains. Chi-stimulated plasmid recombination is dependent on RecBCD but is independent of RecF pathway genes. The distribution of recombination products suggests that high-molecular-weight linear plasmid DNA is a substrate for RecBCD-mediated recombination. Surprisingly, our data also suggest that chi+ plasmids also recombine by the RecBCD pathway in rec+ sbcB+ cells.

Full Text

The Full Text of this article is available as a PDF (536K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bi X, Liu LF. recA-independent and recA-dependent intramolecular plasmid recombination. Differential homology requirement and distance effect. J Mol Biol. 1994 Jan 14;235(2):414–423. [PubMed]
  • Cheng KC, Smith GR. Distribution of Chi-stimulated recombinational exchanges and heteroduplex endpoints in phage lambda. Genetics. 1989 Sep;123(1):5–17. [PMC free article] [PubMed]
  • Cohen A, Clark AJ. Synthesis of linear plasmid multimers in Escherichia coli K-12. J Bacteriol. 1986 Jul;167(1):327–335. [PMC free article] [PubMed]
  • Cohen A, Laban A. Plasmidic recombination in Escherichia coli K-12: the role of recF gene function. Mol Gen Genet. 1983;189(3):471–474. [PubMed]
  • Dabert P, Ehrlich SD, Gruss A. High-molecular-weight linear multimer formation by single-stranded DNA plasmids in Escherichia coli. J Bacteriol. 1992 Jan;174(1):173–178. [PMC free article] [PubMed]
  • Dabert P, Ehrlich SD, Gruss A. Chi sequence protects against RecBCD degradation of DNA in vivo. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12073–12077. [PMC free article] [PubMed]
  • Dianov GL, Kuzminov AV, Mazin AV, Salganik RI. Molecular mechanisms of deletion formation in Escherichia coli plasmids. I. Deletion formation mediated by long direct repeats. Mol Gen Genet. 1991 Aug;228(1-2):153–159. [PubMed]
  • Dixon DA, Kowalczykowski SC. Homologous pairing in vitro stimulated by the recombination hotspot, Chi. Cell. 1991 Jul 26;66(2):361–371. [PubMed]
  • Dixon DA, Kowalczykowski SC. The recombination hotspot chi is a regulatory sequence that acts by attenuating the nuclease activity of the E. coli RecBCD enzyme. Cell. 1993 Apr 9;73(1):87–96. [PubMed]
  • Doherty MJ, Morrison PT, Kolodner R. Genetic recombination of bacterial plasmid DNA. Physical and genetic analysis of the products of plasmid recombination in Escherichia coli. J Mol Biol. 1983 Jul 5;167(3):539–560. [PubMed]
  • Ennis DG, Amundsen SK, Smith GR. Genetic functions promoting homologous recombination in Escherichia coli: a study of inversions in phage lambda. Genetics. 1987 Jan;115(1):11–24. [PMC free article] [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Feinberg AP, Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. [PubMed]
  • Fishel RA, James AA, Kolodner R. recA-independent general genetic recombination of plasmids. Nature. 1981 Nov 12;294(5837):184–186. [PubMed]
  • Fishel RA, Kolodner R. Escherichia coli strains containing mutations in the structural gene for topoisomerase I are recombination deficient. J Bacteriol. 1984 Dec;160(3):1168–1170. [PMC free article] [PubMed]
  • Goldmark PJ, Linn S. Purification and properties of the recBC DNase of Escherichia coli K-12. J Biol Chem. 1972 Mar 25;247(6):1849–1860. [PubMed]
  • James AA, Morrison PT, Kolodner R. Genetic recombination of bacterial plasmid DNA. Analysis of the effect of recombination-deficient mutations on plasmid recombination. J Mol Biol. 1982 Sep 25;160(3):411–430. [PubMed]
  • Karu AE, MacKay V, Goldmark PJ, Linn S. The recBC deoxyribonuclease of Escherichia coli K-12. Substrate specificity and reaction intermediates. J Biol Chem. 1973 Jul 25;248(14):4874–4884. [PubMed]
  • Kleckner N, Bender J, Gottesman S. Uses of transposons with emphasis on Tn10. Methods Enzymol. 1991;204:139–180. [PubMed]
  • Kobayashi I, Murialdo H, Crasemann JM, Stahl MM, Stahl FW. Orientation of cohesive end site cos determines the active orientation of chi sequence in stimulating recA . recBC-mediated recombination in phage lambda lytic infections. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5981–5985. [PMC free article] [PubMed]
  • Kolodner R, Fishel RA, Howard M. Genetic recombination of bacterial plasmid DNA: effect of RecF pathway mutations on plasmid recombination in Escherichia coli. J Bacteriol. 1985 Sep;163(3):1060–1066. [PMC free article] [PubMed]
  • Kusano K, Nakayama K, Nakayama H. Plasmid-mediated lethality and plasmid multimer formation in an Escherichia coli recBC sbcBC mutant. Involvement of RecF recombination pathway genes. J Mol Biol. 1989 Oct 20;209(4):623–634. [PubMed]
  • Lam ST, Stahl MM, McMilin KD, Stahl FW. Rec-mediated recombinational hot spot activity in bacteriophage lambda. II. A mutation which causes hot spot activity. Genetics. 1974 Jul;77(3):425–433. [PMC free article] [PubMed]
  • Lovett ST, Drapkin PT, Sutera VA, Jr, Gluckman-Peskind TJ. A sister-strand exchange mechanism for recA-independent deletion of repeated DNA sequences in Escherichia coli. Genetics. 1993 Nov;135(3):631–642. [PMC free article] [PubMed]
  • Lovett ST, Gluckman TJ, Simon PJ, Sutera VA, Jr, Drapkin PT. Recombination between repeats in Escherichia coli by a recA-independent, proximity-sensitive mechanism. Mol Gen Genet. 1994 Nov 1;245(3):294–300. [PubMed]
  • Luisi-DeLuca C, Lovett ST, Kolodner RD. Genetic and physical analysis of plasmid recombination in recB recC sbcB and recB recC sbcA Escherichia coli K-12 mutants. Genetics. 1989 Jun;122(2):269–278. [PMC free article] [PubMed]
  • Matfield M, Badawi R, Brammar WJ. Rec-dependent and Rec-independent recombination of plasmid-borne duplications in Escherichia coli K12. Mol Gen Genet. 1985;199(3):518–523. [PubMed]
  • Niki H, Ogura T, Hiraga S. Linear multimer formation of plasmid DNA in Escherichia coli hopE (recD) mutants. Mol Gen Genet. 1990 Oct;224(1):1–9. [PubMed]
  • Rose RE. The nucleotide sequence of pACYC184. Nucleic Acids Res. 1988 Jan 11;16(1):355–355. [PMC free article] [PubMed]
  • Simmon VF, Lederberg S. Degradation of bacteriophage lambda deoxyribonucleic acid after restriction by Escherichia coli K-12. J Bacteriol. 1972 Oct;112(1):161–169. [PMC free article] [PubMed]
  • Smith GR. Homologous recombination in procaryotes. Microbiol Rev. 1988 Mar;52(1):1–28. [PMC free article] [PubMed]
  • Taylor AF, Smith GR. Substrate specificity of the DNA unwinding activity of the RecBC enzyme of Escherichia coli. J Mol Biol. 1985 Sep 20;185(2):431–443. [PubMed]
  • Watson N. A new revision of the sequence of plasmid pBR322. Gene. 1988 Oct 30;70(2):399–403. [PubMed]
  • Willetts NS, Clark AJ. Characteristics of some multiply recombination-deficient strains of Escherichia coli. J Bacteriol. 1969 Oct;100(1):231–239. [PMC free article] [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]
  • Zaman MM, Boles TC. Chi-dependent formation of linear plasmid DNA in exonuclease-deficient recBCD+ strains of Escherichia coli. J Bacteriol. 1994 Aug;176(16):5093–5100. [PMC free article] [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...