Summary
We have isolated a mutant H group plasmid temperature-sensitive for plasmid maintenance. Unlike the wild type plasmid (pSD114), the mutant (pDT4) was eliminated at 37° C and also at 30° C after novobiocin treatment. The mutant plasmid interfered with host cell growth at the non-permissive temperature. Conjugative transfer of the mutant was reduced at 30° C compared to the wild-type plasmid. Introduction of a coumermycin-novobiocin resistance DNA gyrase (cou) mutation into Escherichia coli prevented pDT4 elimination by novobiocin but did not affect the temperature-sensitive phenotype. The evidence indicates that the mutant plasmid used bacterial DNA gyrase for replication. Models to account for the behaviour of this unusual mutant are discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Anderson, E.S.: The problem and implications of chloramphenicol resistance in the typhoid bacillus. J. Hyg. (Camb.) 74, 289–299 (1975)
Bachmann, B.: Pedigress of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 36, 525–557 (1972)
Cuzin, F., Jacob, F.: Mutations de l'épisome F d'Escherichia coli K12 II. Mutants à réplications thermosensible. Ann. Inst. Pasteur 112, 398–418 (1967)
Gellert, M., O'Dea, M.H., Itoh, T., Tomizawa, J.-I.: Novobiocin and coumermycin inhibit DNA supercoiling catalyzed by DNA gyrase. Proc. Natl. Acad. Sci. (U.S.A.) 73, 4474–4478 (1976)
Grant, R.B., Di Mambro, L.: Antimicrobial resistance and resistance plasmids in Salmonella from Ontario, Canada. Can. J. Microbiol. 23, 1266–1273 (1977)
Grindley, N.D.F., Humphreys, G.O., Anderson, E.S.: Molecular studies of R factor compatibility groups. J. Bacteriol. 115, 387–398 (1973)
Hansen, J.B., Olsen, R.H.: Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J. Bacteriol. 135, 227–238 (1978)
Hashimoto-Gotoh, T., Sekiguchi, M.: Mutations to temperaturesensitivity in R plasmid pSC101. J. Bacteriol. 131, 405–412 (1977)
Higgins, N.P., Peeples, C.L., Sugino, A., Cozzarelli, N.R.: Purification of subunits of Escherichia coli DNA gyrase and reconstitution of enzymatic activity. Proc. Natl. Acad. Sci. (U.S.A.) 75, 1773–1777 (1978)
Itoh, T., Tomizawa, J.-I.: Involvement of DNA gyrase in bacteriophage T7 DNA replication. Nature 270, 78–80 (1977)
Jacob, A., Shapiro, J.A., Yamamoto, L., Smith, D.I., Cohen, S.N., Berg, D.: Plasmids studied in Escherichia coli and other enteric bacteria. In: DNA insertion elements, plasmids and episomes (A.I. Bukhari, J.A. Shapiro and S.L. Adhya, eds.), pp. 607–638 Cold Spring Harbor, New York: Cold Spring Harbor Laboratory 1977
Jaffé-Brachet, A., D'Ari, R.: Maintenance of bacteriophage P1 plasmid. J. Virol. 23, 476–482 (1977)
Kingsbury, D.T., Helinski, D.R.: Temperature-sensitive mutants for the replication of plasmids in Escherichia coli 1. Isolation and specificity of host and plasmid mutations. Genetics 74, 17–31 (1973)
Kingsbury, D.T., Sieckmann, D.G., Helinski, D.R.: Temperature-sensitive mutants for the replication of plasmids in Escherichia coli. II Properties of host and plasmid mutations. Genetics 74, 1–16 (1973)
Koyama, A.H., Yura, T.: Plasmid mutations affecting self-maintenance and host growth in Escherichia coli. J. Bacteriol. 122, 80–88 (1975)
Kretchmer, P.J., Chang, A.C.Y., Cohen, S.N.: Indirect detection of bacterial plasmid lacking identifiable phenotypic properties. J. Bacteriol. 124, 225–231 (1975)
McHugh, G.L., Swartz, M.N.: Elimination of plasmids from several bacterial species by novobiocin. Antimicrob. Agents Chemother. 12, 423–426 (1977)
Meyers, J.A., Sanchez, D., Elwell, L.P., Falkow, S.: Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J. Bacteriol. 127, 1529–1537 (1976)
Miller, J.H.: Experiments in molecular genetics. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory 1972
Mizuuchi, K., O'Dea, M.H., Gellert, M.: DNA gyrase: subunit structure and ATPase activity of the purified enzyme. Proc. Natl. Acad. Sci. U.S.A. 75, 5960–5963 (1978)
Scott, J.R.: Genetic studies on bacteriophage P1. Virology 36, 564–573 (1968)
Smith, H.W.: Thermosensitive transfer factors in chloramphenicolresistant strains of Salmonella typhi. Lancet 11, 281 (1974)
Smith, H.W., Parsell, Z., Green, P.: Thermosensitive antibiotic resistance plasmids in enterobacteria. J. Gen. Microbiol. 109, 37–47 (1978)
Stadenbauer, W.L.: Replication of the ampicillin resistance plasmid RSF1030 in extracts of Escherichia coli: Separation of the replicative cycle into early and late stages. Mol. Gen. Genet. 156, 27–34 (1977)
Sugino, A., Higgins, N.P., Brown, P.O., Peebles, C.L., Cozzarelli, N.R.: Energy coupling in DNA gyrase and mechanism of action of novobiocin. Proc. Natl. Acad. Sci. (U.S.A.) 75, 4838–4842 (1978)
Taylor, D.E., Grant, R.B.: Incompatibility and bacteriophage inhibition properties of N-1, a plasmid belonging to the H2 incompatibility group. Mol. Gen. Genet. 153, 5–10 (1977a)
Taylor, D.E., Grant, R.B.: R plasmids of the S incompatibility group belong to the H2 incompatibility group. Antimicrob. Agents Chemother. 12, 431–434 (1977b)
Taylor, D.E., Shermer, M., Grant, R.B.: Incidence of the H2 group of plasmids in chloramphenicol-sensitive salmonella isolated in 1974 from clinical sources in Ontario. Can. J. Microbiol. 24, 600–607 (1978)
Taylor, D.E., Summers, A.O.: Association of tellurium resistance and bacteriophage inhibition conferred by R plasmids. J. Bacteriol. 137, 1430–1433 (1979)
Timoney, J.F.: The epidemiology and genetics of antibiotic resistance of Salmonella typhimurium isolated from diseased animals in New York. J. Infect. Dis. 137, 67–73 (1978)
Yanofsky, C., Ito, J., Horn, V.: Amino acid replacements and the genetic code. Cold Spring Harbor Symp. Quant. Biol. 31, 151–162 (1966)
Author information
Authors and Affiliations
Additional information
Communicated by W. Arber
Rights and permissions
About this article
Cite this article
Taylor, D.E., Levine, J.G. Characterization of a plasmid mutation affecting maintenance, transfer and elimination by novobiocin. Molec. Gen. Genet. 174, 127–133 (1979). https://doi.org/10.1007/BF00268350
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00268350