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Loss of Hfr DNA from Escherichia coli merozygotes during inhibition of conjugation by nalidixic acid

Published online by Cambridge University Press:  14 April 2009

Robert G. Lloyd ,
Janet Hart  and
Sandra Johnson
Robert G. Lloyd
Affiliation:
Department of Genetics, University of Nottingham, Nottingham NG7 2RD, England
Janet Hart
Affiliation:
Department of Genetics, University of Nottingham, Nottingham NG7 2RD, England
Sandra Johnson
Affiliation:
Department of Genetics, University of Nottingham, Nottingham NG7 2RD, England
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The effect of nalidixic acid on conjugal recombination was studied in matings with recipient strains carrying recA200, a mutation which confers a thermosensitive Rec− phenotype. Addition of nalidixic acid to Hfr Nals × F−NalRrecA200 matings at low temperature (35 °C) caused a sharp 10- to 20-fold decline in the yield of recombinants if plating on selective agar was delayed. Two separate processes were identified as being responsible for this decline. Those merozygotes in which the transferred DNA was free of the donor cell lost the ability to form recombinants through inactivation of this DNA, an effect which could be prevented by using exonuclease deficient (recB sbcB) recipients or by prior growth of exonuclease proficient recipients in medium containing 0·25 M sodium chloride. No more than 50% of the observed loss of recombinants could be attributed to this effect. The remaining merozygotes lost their ability for recombinant formation provided mating pairs, and presumably the displaced donor DNA strand, remained intact. This process was thought to involve withdrawal of transferred DNA (DeHaan & Gross, 1962) and was studied in isolation in matings with recA+, or recA200 recB sbcB recipients. A mechanism involving re-annealing of the displaced Hfr DNA to the donor molecule as a result of nalidixic inhibition of gyrase activity in the donor causing relaxation of DNA supercoils is proposed to account for this withdrawal event.

Type
Research Article
Information
Genetics Research , Volume 36 , Issue 1 , August 1980 , pp. 69 - 79
Copyright
Copyright © Cambridge University Press 1980

References

REFERENCES

Bachmann, B. J.Low, K. B. & Taylor, A. L. (1976). Recalibrated linkage map of Escherichia coli K-12. Bacteriological Reviews 40, 116–167.CrossRefGoogle ScholarPubMed
Barbour, S. D. (1967). Effect of nalidixic acid on conjugational transfer and expression of episomal Lac genes in Escherichia coli K-12. Journal of Molecular Biology 28, 373–376.CrossRefGoogle Scholar
Bouck, N. & Adelberg, E. A. (1970). Mechanism of action of nalidixic acid on conjugating bacteria. Journal of Bacteriology 102, 688–701.CrossRefGoogle ScholarPubMed
DeHaan, P. G. & Gross, J. D. (1962). Transfer delay and chromosome withdrawal during conjugation in Escherichia coli. Genetical Research 3, 251–272.CrossRefGoogle Scholar
Demerec, M., Adelberg, E. A., Clark, A. J. & Hartman, P. E. (1966). A proposal for a uniform nomenclature in bacterial genetics. Genetics 54, 61–74.CrossRefGoogle ScholarPubMed
Drlica, K. & Snyder, M. (1978). Superhelical Escherichia coli DNA: relaxation by coumermycin. Journal of Molecular Biology 120, 145–154.CrossRefGoogle ScholarPubMed
Gellert, M., Mizuuchi, K., O'Dea, M. H., Itoh, T. & Tomizawa, J. (1977). Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proceedings of the National Academy of Sciences, U.S.A. 74, 4772–4776.CrossRefGoogle ScholarPubMed
Hane, M. W. (1971). Some effects of nalidixic acid on conjugation in Escherichia coli K-12. Journal of Bacteriology 105, 46–56.CrossRefGoogle ScholarPubMed
Hane, M. W. & Wood, T. H. (1969). Escherichia coli K-12 mutants resistant to nalidixic acid: genetic mapping and dominance studies. Journal of Bacteriology 99, 238–241.CrossRefGoogle ScholarPubMed
Itoh, T. & Tomizawa, J. (1971). Inactivation of chromosomal fragments transferred from Hfr strains. Genetics 68, 1–11.CrossRefGoogle ScholarPubMed
Kingsman, A. & Willetts, N. (1978). The requirements for conjugal DNA synthesis in the donor strain during F lac transfer. Journal of Molecular Biology 122, 287–300.CrossRefGoogle Scholar
Kreuzer, K. N., McEntee, K., Geballe, A. P. & Cozzarelli, N. R. (1978). Lambda transducing phages for the nalA gene of Escherichia coli and conditional lethal nalA mutations. Molecular and General Genetics 167, 129–137.CrossRefGoogle ScholarPubMed
Kushner, S. R., Nagaishi, H. & Clark, A. J. (1972). Indirect suppression of recB and recC mutations by exonuclease I deficiency. Proceedings of the National Academy of Sciences, U.S.A. 69, 1366–1370.CrossRefGoogle ScholarPubMed
Lloyd, R. G. & Johnson, S. (1979). Kinetics of recA function in conjugational recombinant formation. Molecular and General Genetics 169, 219–228.CrossRefGoogle ScholarPubMed
Lloyd, R. G. & Low, B. (1976). Some genetic consequences of changes in the level of recA gene function in Escherichia coli K-12. Genetics 84, 675–695.CrossRefGoogle ScholarPubMed
Lloyd, R. G., Low, B., Godson, G. N. & Birge, E. A. (1974). Isolation and characterization of an Escherichia coli K-12 mutant with a temperature-sensitive RecA− phenotype. Journal of Bacteriology 120, 407–415.CrossRefGoogle ScholarPubMed
Low, K. B. (1973). Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. Journal of Bacteriology 113, 798–812.CrossRefGoogle ScholarPubMed
Low, K. B. & Wood, T. H. (1965). A quick and efficient method for interruption of bacterial conjugation. Genetical Research 6, 300–303.CrossRefGoogle ScholarPubMed
Mizuuchi, K., O'Dea, M. H. & Gellert, M. (1978). DNA gyrase: subunit structure and ATPase activity of the purified enzyme. Proceedings of the National Academy of Sciences, U.S.A. 75, 5960–5963.CrossRefGoogle ScholarPubMed
Rupp, W. D. & Ihler, G. (1968). Strand selection during bacterial mating. Cold Spring Harbor Symposia on Quantitative Biology 33, 647–650.CrossRefGoogle ScholarPubMed
Sarathy, P. V. & Siddiqi, O. (1973). DNA synthesis during bacterial conjugation. II. Is DNA replication in the Hfr obligatory for chromosome transfer? Journal of Molecular Biology 78, 443–451.CrossRefGoogle ScholarPubMed
Shibata, T., Dasgupta, C., Cunningham, R. P. & Radding, C. M. (1979). Purified Escherichia coli recA protein catalyses homologous pairing of superhelical DNA and single-stranded fragments. Proceedings of the National Academy of Sciences, U.S.A. 76, 1638–1642.CrossRefGoogle Scholar
Siddiqi, O. & Fox, M. S. (1973). Integration of donor DNA in bacterial conjugation. Journal of Molecular Biology 77, 101–123.CrossRefGoogle ScholarPubMed
Sugino, A., Peebles, C. L., Kreuzer, K. N. & Cozzarelli, N. R. (1977). Mechanism of action of nalidixic acid: purification of Escherichia coli nalA gene product and its relationship to DNA gyrase and a novel nicking-closing enzyme. Proceedings of the National Academy of Sciences, U.S.A. 74, 4767–4771.CrossRefGoogle Scholar