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Accumulation of replicating bacterial plasmid DNA during thymine limitation or hydroxyurea treatment

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Summary

The rate of DNA chain elongation of a thymine auxotroph of Proteus mirabilis harboring an R factor was reduced by either thymine limitation or treatment with hydroxyurea. Using CsCl density gradient centrifugation, sedimentation in sucrose gradients, and electron microscopy, it has been shown that the fraction of total R factor DNA which is in the process of replication is greatly increased when the rate of DNA chain elongation is reduced while the initiation of plasmid replication continues at the normal rate. This makes possible the isolation and characterization of replicating plasmid DNA. Electron microscopy revealed that replicating R factor DNA consists of double-branched (theta type) circular molecules.

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References

  • Barnes, M. H., Rownd, R.: Effect of thymine limitation on chromosomal deoxyribonucleic acid synthesis in Proteus mirabilis. J. Bact. 111, 750–757 (1972)

    Google Scholar 

  • Bird, R. E., Louarn, J., Martuscelli, J., Caro, L.: Origin and sequence of chromosome replication in Escherichia coli. J. molec. Biol. 70, 549–566 (1972)

    Google Scholar 

  • Cairns, J.: The chromosome of Escherichia coli. Cold Spr. Harb. Symp. quant. Biol. 28, 43–46 (1963)

    Google Scholar 

  • Cooper, S., Helmstetter, C.: Chromosome replication and the division cycle of Escherichia coli B/r. J. molec. Biol. 31, 519–540 (1968)

    Google Scholar 

  • Dressler, D., Wolfson, J., Magazin, M.: Initiation and reinitiation of DNA synthesis during replication of bacteriophage T7. Proc. nat. Acad. Sci. (Wash.) 69, 998–1002 (1972)

    Google Scholar 

  • Freifelder, D.: Molecular weights of coliphages and coliphage DNA. J. molec. Biol. 54, 567–577 (1970)

    Google Scholar 

  • Enman, R., Schnös, M.: Partial denaturation of thymine- and 5-bromouracil-containing λ DNA in alkali. J. molec. Biol. 49, 93–98 (1970)

    Google Scholar 

  • Kasamatsu, H., Rownd, R.: Replication of R-factors in Proteus mirabilis: Replication under relaxed control. J. molec. Biol. 51, 473–489 (1970)

    Google Scholar 

  • Masters, M., Broda, P.: Evidence for the bidirectional replication of the Escherichia coli chromosome. Nature (Lond.) New Biol. 232, 137–140 (1971)

    Google Scholar 

  • Nakaya, R., Nakamura, A., Murata, Y.: Resistance transfer agents in Shigella. Biochem. biophys. Res. Commun. 31, 654–659 (1960)

    Google Scholar 

  • Neuhard, J.: Studies on the acid-soluble nucleotide pool in E. coli. IV. Effects of hydroxyrea. Biochim. biophys. Acta (Amst.) 145, 1–6 (1967)

    Google Scholar 

  • O'Donovan, G., Neuhard, J.: Pyrimidine metabolism in microorganisms. Bact. Rev. 34, 278–343 (1970)

    Google Scholar 

  • Perlman, D., Rownd, R.: Transition of the R factor NR1 in Proteus mirabilis: Molecular structure and replication of NR1 DNA. Submitted to J. Bact. (1975)

  • Prescott, D. M., Kuempel, P. L.: Bidirectional replication of the chromosome in Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 69, 2842–2845 (1972)

    Google Scholar 

  • Pritchard, R., Zaritsky, A.: Effect of thymine concentration on the replication velocity of DNA in a thymineless mutant of Escherichia coli. Nature (Lond.) 226, 126–131 (1970)

    Google Scholar 

  • Rownd, R.: Replication of a bacterial episome under relaxed control. J. molec. Biol. 44, 387–402 (1969)

    Google Scholar 

  • Rownd, R.: Initiator proteins for R-factor replication. Fed. Proc. 30, 1313 (1971)

    Google Scholar 

  • Rownd, R.: Mickel, S.: Dissociation and reassociation of the RTF and r-determinants of the R-factor N R1 in Proteus mirabilis. Nature (Lond.) New Biol. 234, 40–43 (1971)

    Google Scholar 

  • Rownd, R., Nakaya, R., Nakamura, A.: Molecular nature of the drug-resistance factors of the enterobacteriaceae. J. molec. Biol. 17, 376–393 (1966)

    Google Scholar 

  • Schnös, M., Inman, R. B.: Position of branch points in replicating λ DNA. J. molec. Biol. 51, 61–73 (1970)

    Google Scholar 

  • Schnös, M., Inman, R. B.: Starting point and direction of replication in P2 DNA. J. molec. Biol. 55, 31–38 (1971)

    Google Scholar 

  • Sueoka, N., Quinn, W.: Membrane attachment of the chromosome replication origin in Bacillus subtilis. Cold Spr. Harb. Symp. quant. Biol. 33, 695–705 (1968)

    Google Scholar 

  • Wake, R. G.: Circularity of the Bacillus subtilis chromosome and further studies on its bidirectional replication. J. molec. Biol. 77, 569–575 (1973)

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Molecular Biology, University of Wisconsin, Madison, Wisconsin

    Dan Perlman & Robert H. Rownd

  2. Department of Biochemistry, University of Wisconsin, Madison, Wisconsin

    Dan Perlman & Robert H. Rownd

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  1. Dan Perlman
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  2. Robert H. Rownd
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Communicated by W. Maas

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Perlman, D., Rownd, R.H. Accumulation of replicating bacterial plasmid DNA during thymine limitation or hydroxyurea treatment. Molec. Gen. Genet. 138, 281–291 (1975). https://doi.org/10.1007/BF00264797

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  • DOI: https://doi.org/10.1007/BF00264797

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