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Linear multimer formation of plasmid DNA in Escherichia coli hopE (recD) mutants

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Summary

The hopE mutants of Escherichia coli, which cannot stably maintain a mini-F plasmid during cell division, have mutations in the recD gene coding for subunit D of the RecBCD enzyme (exonuclease V). A large amount of linear multimer DNA of mini-F and pBR322 plasmids accumulates in these hopE mutants. The linear multimers of plasmid DNA in the hopE (recD) mutants accumulate in sbc + genetic backgrounds and this depends on the recA + gene function. Linear plasmid multimers also accumulated in a recBC xthA triple mutant, but not an isogenic xthA mutant or an isogenic recBC mutant. The recBC xthA mutant is defective in the conjugative type of recombination. Linear plasmid multimers were not detected in the recBC strain. We propose models to account for linear multimer formation of plasmids in various mutants.

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References

  • Amundsen SK, Taylor AF, Chaudhury AM, Smith GR (1986) recD: the gene for an essential third subunit of exonuclease V. Proc Natl Acad Sci USA 83:5558–5562

    Google Scholar 

  • Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557

    Google Scholar 

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

    Google Scholar 

  • Barbour SD, Clark AJ (1970) Biochemical and genetic studies of recombination proficiency in Escherichia coli. Enzymatic activity associated with the recB + and recC + genes. Proc Natl Acad Sci USA 65:955–961

    Google Scholar 

  • Bassett CL, Kushner SR (1984) Exonuclease I, III, and V are required for stability of ColE1-related plasmid in Escherichia coli. J Bacteriol 157:661–664

    Google Scholar 

  • Biek DP, Cohen SN (1986) Identification and characterization of recD, a gene affecting plasmid maintenance and recombination in Escherichia coli. J Bacteriol 167:594–603

    Google Scholar 

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

    Google Scholar 

  • Brutlag D, Kornberg A (1972) Enzymatic synthesis of deoxyribonucleic acid XXXVI. A proofreading function for the 3′–5′ exonuclease activity in deoxyribonucleic acid polymerases. J Biol Chem 247:241–248

    Google Scholar 

  • Buttin G, Wright M (1968) Enzymatic DNA degradation in E. coli: its relationship to synthetic processes at the chromosomal level. Cold Spring Harbor Symp Quant Biol 33:259–269

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15 cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Chaudhury AM, Smith GR (1984) A new class of Escherichia coli recBC mutants: implications for the role of RecBC enzyme in homologous recombination. Proc Natl Acad Sci USA 81:7850–7854

    Google Scholar 

  • Clark AJ (1973) Recombination deficient mutants of E. coli and other bacteria. Annu Rev Genet 7:67–86

    Google Scholar 

  • Cohen A, Clark AK (1986) Synthesis of linear plasmid multimers in Escherichia coli K-12. J Bacteriol 167:327–335

    Google Scholar 

  • Finch PW, Storey A, Brown K, Hickson ID, Emmerson PT (1986) Complete nucleotide sequence of recD, the structural gene for the a subunit of exonuclease V of Escherichia coli. Nucleic Acids Res 14:8583–8594

    Google Scholar 

  • Frischauf AM, Lehrach H, Poustka A, Murray N (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842

    Google Scholar 

  • Gillen JR, Karu AE, Nagaishi H, Clark AJ (1977) Characterization of the deoxyribonuclease determined by lambda reverse as exonuclease VIII of Escherichia coli. J Mol Biol 113:27–41

    Google Scholar 

  • Irino N, Nakayama K, Nakayama H (1986) The recQ gene of Escherichia coli K12: primary structure and evidence for SOS regulation. Mol Gen Genet 205:298–304

    Google Scholar 

  • Ish-Horowicz D, Burke JF (1981) Rapid and efficient cosmid cloning. Nuleic Acids Res 9:2989–2998

    Google Scholar 

  • Joseph JW, Kolodner R (1983) Exonuclease VIII of Escherichia coli. II. Mechanism of the action. J Biol Chem 258:10418–10424

    Google Scholar 

  • Kolodner R, Fishel RA, Howard M (1985) Genetic recombination of bacterial plasmid DNA: effect of RecF pathway mutations on plasmid recombination in Escherichia coli. J Bacteriol 163:1060–1066

    Google Scholar 

  • Konforti BB, Davis RW (1987) 3′ homologous free ends are required for stable joint molecule formation by the RecA and single-stranded binding proteins of Escherichia coli. Proc Natl Acad Sci USA 84:690–694

    Google Scholar 

  • Kornberg A (1980) DNA replication. WH Freeman and Company, San Francisco, pp 323–324

    Google Scholar 

  • Kushner SR, Nagaishi H, Clark AJ (1974) Isolation of exonuclease VIII: the enzyme associated with the sbcA in direct suppressor. Proc Natl Acad Sci USA 71:3593–3597

    Google Scholar 

  • Lloyd RG, Buckman C (1985) Identification and analysis of sbcC mutations in commonly used recBC sbcB strains of Escherichia coli K12. J Bacteriol 164:836–844

    Google Scholar 

  • Lloyd RG, Porten MC, Buckman C (1988) Effect of recF, recJ, recN, recO and ruv mutations on ultraviolet survival and genetic recombination in a recD strain of Escherichia coli K12. Mol Gen Genet 212:317–324

    Google Scholar 

  • Lovett ST, Clark AJ (1983) Genetic analysis of regulation of the recF pathway of recombination in Escherichia coli K-12. J Bacteriol 153:1471–1478

    Google Scholar 

  • Lovett ST, Luisi-Deluca C, Kolodner RD (1988) The genetic dependence of recombination in recD mutants of Escherichia coli. Genetics 120:37–46

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Miki T, Chang Z-T, Horiuchi T (1984) Control of cell division by sex factor F in Escherichia coli. J Mol Biol 174:627–646

    Google Scholar 

  • Nakayama H, Nakayama K, Nakayama R, Nakayama Y (1982) Recombination-deficient mutations and thymineless death in Escherichia coli: reciprocal effects of recBC and recF and indifference of recA mutations. Can J Microbiol 28:425–430

    Google Scholar 

  • Niki H, Ichinose C, Ogura T, Mori H, Morita M, Hasegawa M, Kusukawa N, Hiraga S (1988) Chromosomal genes essential for stable maintenance of the mini-F plasmid in Escherichia coli. J Bacteriol 171:1496–1505

    Google Scholar 

  • O'Connor MB, Kilbane JJ, Malamy MH (1986) Site-specific and illegitimate recombination in the oriV1 region of the F factor. J Mol Biol 189:85–102

    Google Scholar 

  • Ogura T, Hiraga S (1983) Mini-F plasmid genes that couple host cell division to plasmid proliferation. Proc Natl Acad Sci USA 80:4784–4788

    Google Scholar 

  • Oishi M (1969) An ATP-dependent deoxyribonuclease from Escherichia coli with a possible role in genetic recombination. Proc Natl Acad Sci USA 64:1292–1299

    Google Scholar 

  • Projan SJ, Carleton S, Novick RP (1983) Determination of plasmid copy number by fluorescence densitometry. Plasmid 9:182–190

    Google Scholar 

  • Rosamond JW, Telander KM, Linn S (1979) Modulation of the RecBC enzyme of Escherichia coli Kl2 by Ca++. J Biol Chem 254:8646–8652

    Google Scholar 

  • Rosenberg SM (1987) Chi-stimulated patches are heteroduplex, with recombinant information on the phage λ, r chain. Cell 48:855–865

    Google Scholar 

  • Sanger F, Coulson AR (1975) A rapid method for determining sequence in DNA by primed synthesis with DNA polymerase. J Mol Biol 94:441–448

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sasaki M, Fujiyoshi T, Shimada K, Takagi Y (1982) Fine structure of the recB and recC region of Escherichia coli. Biochem Biophys Res Commun 109:414–422

    Google Scholar 

  • Seelke P, Kline B, Aleff R, Porter RD, Shields NS (1987) Mutations in the recD gene of Escherichia coli that raise the copy number of certain plasmids. J Bacteriol 169:4841–4844

    Google Scholar 

  • Silberstein Z, Cohen A (1987) Synthesis of linear multimers of oriC and pBR322 derivatives in Escherichia coli K-12: Role of recombination and replication functions. J Bacteriol 169:131–137

    Google Scholar 

  • Silhavy TJ, Berman ML, Enquist LW (1984) Experiments with gene fusions. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 137–139

    Google Scholar 

  • Smith GW (1988) Homologous recombination in procaryotes. Microbiol Rev 52:1–28

    Google Scholar 

  • White BJ, Hochhauser SJ, Cintón NM, Weiss B (1976) Genetic mapping of xthA, the structural gene of exonuclease III in Escherichia coli K-12. J Bacteriol 126:1082–1088

    Google Scholar 

  • Wright M, Buttin G, Hurwitz J (1971) The isolation and characterization from Escherichia coli of an adenosine triphosphate-dependent deoxyribonuclease directed by recB, C genes. J Biol Chem 246:6543–6555

    Google Scholar 

  • Yajko DM, Weiss B (1975) Mutations simultaneously affecting endonuclease 11 and exonuclease III in Escherichia coli. Proc Natl Acad Sci USA 72:688–692

    Google Scholar 

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Authors and Affiliations

  1. Department of Molecular Genetics, Institute for Medical Genetics, Kumamoto University Medical School, 862, Kumamoto, Japan

    Hironori Niki, Teru Ogura & Sota Hiraga

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  1. Hironori Niki
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  2. Teru Ogura
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  3. Sota Hiraga
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Communicated by M. Sekiguchi

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Niki, H., Ogura, T. & Hiraga, S. Linear multimer formation of plasmid DNA in Escherichia coli hopE (recD) mutants. Molec. Gen. Genet. 224, 1–9 (1990). https://doi.org/10.1007/BF00259444

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  • DOI: https://doi.org/10.1007/BF00259444

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