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Isolation and characterization of DNA topoisomerase II from cauliflower inflorescences

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Summary

Type II DNA topoisomerase has been isolated from inflorescences of cauliflower (Brassica oleracea var. botrytis) through a sequence of polyethylene glycol fractionation, ammonium sulfate precipitation, and column chromatography on CM-Sephadex, hydroxyapatite and phosphocellulose. The molecular weight of the native enzyme, based on sedimentation coefficient (9S) and gel filtration analysis (Stokes radius, 60 Å), was estimated to be 223 000. This enzyme was able to catalyze fully the relaxation of supercoiled DNA by breaking and then rejoining the double-stranded DNA. The breaking reaction was reversible by a change in salt concentrations. When an antitumor drug, 4′-(9-acridinylamino)-methanesulfon-m-anisidide, was added to the topoisomerase reaction, DNA cleavage fragments were accumulated; and this suggested that the drug interfered with the reaction at the rejoining step. This enzyme also catalyzed the formation of DNA catenanes in the presence of 8% polyethylene glycol or histone H1, while few catenanes were formed in the presence of spermidine, which was highly effective on a bacterial enzyme.

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References

  1. Baldi MI, Benedetti P, Mattoccia E, Tocchini-Valentini GP: In vitro catenation and decatenation of DNA and a novel eucaryotic ATP-dependent topoisomerase. Cell 20:461–467, 1980.

    Google Scholar 

  2. Cozzarelli NR: DNA topoisomerases. Cell 22:327–328, 1980.

    Google Scholar 

  3. DiNardo S, Voelkel K, Sternglanz R: DNA topoisomerase II mutant of Saccharomyces cerevisiae: Topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication. Proc Natl Acad Sci USA 81:2612–2620, 1984.

    Google Scholar 

  4. Dynan WS, Jendrisak JJ, Hager DA, Burgess RR: Purification and characterization of wheat germ DNA topoisomerase I (nicking-closing enzyme). J Biol Chem 256:5860–5865, 1981.

    Google Scholar 

  5. Fukasawa H, Hamada Y: Properties of soluble chromatins isolated from apical and axial parts of cauliflower inflorescence. Exp Cell Res 88:352–358, 1974.

    Google Scholar 

  6. Fukata H, Fukasawa H: Isolation and partial characterization of two distinct DNA topoisomerases from cauliflower inflorescence. J Biochem 91:1337–1342, 1982.

    Google Scholar 

  7. Fukata H, Fukasawa H: Further investigations on the characterization of DNA topoisomerases isolated from cauliflower inflorescence. Jpn J Genet 58:425–432, 1983.

    Google Scholar 

  8. Fukata H, Fukasawa H: Enzyme activity and processivity of DNA relaxation of salt concentrations of cauliflower DNA topoisomerase II. Jpn J Genet 59:303–306, 1984.

    Google Scholar 

  9. Gellert M: DNA topoisomerases. Ann Rev Biochem 50:879–910, 1981.

    Google Scholar 

  10. Goto T, Wang JC: Yeast DNA topoisomerase II. An ATP-dependent type II topoisomerase that catalyzes the catenation, decatenation, unknotting, and relaxation of double-stranded DNA rings. J Biol Chem 257:5866–5872, 1982.

    Google Scholar 

  11. Higgins NP, Peebles CL, Sugino A, Cozzarelli NR: Purification of subunits of Escherichia coli DNA gyrase and reconstitution of enzymatic activity. Proc Natl Acad Sci USA 75:1773–1777, 1978.

    Google Scholar 

  12. Hsieh T, Brutlag D: ATP-dependent DNA topoisomerase from D. melanogaster reversibly catenates duplex DNA rings. Cell 21:115–125, 1980.

    Google Scholar 

  13. Hsieh T, Wang JC: Thermodynamic properties of superhelical DNAs. Biochem 14:527–535, 1975.

    Google Scholar 

  14. Kaguni JM, Kornberg A: Topoisomerase I confers specificity in enzymatic replication of the E. coli chromosome origin. Cell 38:183–190, 1984.

    Google Scholar 

  15. Kikuchi Y, Nash H: Integrative recombination of bacteriophage λ: Requirement for supertwisted DNA in vivo and characterization of int. Cold Spring Harbor Symp Quant Biol 43:1099–1109, 1978.

    Google Scholar 

  16. Krasnow MA, Cozzarelli NR: Catenation of DNA rings by topoisomerases. Mechanism of control by spermidine. J Biol Chem 257:2687–2693, 1982.

    Google Scholar 

  17. Kreuzer KN, Cozzarelli NR: Formation and resolution of DNA catenanes by DNA gyrase. Cell 20:245–254, 1980.

    Google Scholar 

  18. Lee CH, Mizushima H, Kakefuda T: Unwinding of double-stranded DNA helix by dehydration. Proc Natl Acad Sci USA 78:2838–2842, 1981.

    Google Scholar 

  19. Liu LF, Rowe TC, Yang L, Tewey KM, Chen GL: Cleavage of DNA by mammalian DNA topoisomerase II. J Biol Chem 258:15365–15370, 1983.

    Google Scholar 

  20. Liu LF, Liu CC, Alberts BM: Type II DNA topoisomerase: Enzymes that can unknot a topologically knotted DNA molecule via a reversible double-strand break. Cell 19: 697–707, 1981.

    Google Scholar 

  21. Low RL, Kaguni JM, Kornberg A: Potent catenation of supercoiled and gapped DNA circles by topoisomerase I in the presence of a hydrophilic polymer. J Biol Chem 259:4576–4581, 1984.

    Google Scholar 

  22. Marini JC, Miller KG, Englund PT: Decatenation of kinetoplast DNA by topoisomerases. J Biol Chem 255:4976–4979, 1980.

    Google Scholar 

  23. Mattern MR, Painter RB: Dependence of mammalian DNA replication on DNA supercoiling. II. Effects of novobiocin on DNA synthesis in chinese hamster ovary cells. Biochim Biophys Acta 563:306–312, 1979.

    Google Scholar 

  24. Miller KG, Liu LF, Englund PT: A homologeneous type II DNA topoisomerase from HeLa cell nuclei. J Biol Chem 256:9334–9339, 1981.

    Google Scholar 

  25. Mizuuchi K, Fisher LM, O'Dea MH, Gellert M: DNA gyrase action involves the introduction of transient double-stranded breaks into DNA. Proc Natl Acad Sci USA 77:1847–1851, 1980.

    Google Scholar 

  26. Nelson EM, Tewey KM, Liu LF: Mechanism of antitumor drug action: Poisoning of mammalian DNA topoisomerase II on DNA by 4′-(9-acridinylamino)-methanesulfon-m anisidide. Proc Natl Acad Sci USA 81:1361–1365, 1984.

    Google Scholar 

  27. Nishio A, Uyeki EM: Inhibition of DNA synthesis in permeabilized L cells by novobiocin. Biochem Biophys Res Commun 106:1448–1455, 1982.

    Google Scholar 

  28. Riou GF, Gabillot M, Barrois M, Breitburd F, Orth G: A type II-DNA topoisomerase and a catenating protein from the transplantable VX2 carcinoma. Eur J Biochem 146:483–488, 1985.

    Google Scholar 

  29. Sander M, Hsieh T: Double strand DNA cleavage by type II DNA topoisomerase from Drosophila melanogaster. J Biol Chem 258:8421–8428, 1983.

    Google Scholar 

  30. Shelton ER, Osheroff N, Brutlag DL: DNA topoisomerase II from Drosophila melanogaster. Purification and physical characterization. J Biol Chem 258:9530–9535, 1983.

    Google Scholar 

  31. Shure M, Pulleyblank DE, Vinograd J: The problems of of eukaryotic and prokaryotic DNA packaging and in vivo conformation posed by superhelix density heterogeneity. Nucl Acids Res 4:1183–1205, 1977.

    Google Scholar 

  32. Siedlecki J, Zimmermann W, Weissbach A: Characterization of a prokaryotic topoisomerase I activity in chloroplast extracts from spinach. Nucl Acids Res 11:1523–1536, 1983.

    Google Scholar 

  33. Siegel LM, Monty KJ: Determination of molecular weight and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfide and hydroxylamine reductases. Biochim Biophys Acta 112:346–362, 1966.

    Google Scholar 

  34. Steck T, Drlica K: Bacterial chromosome segregation: Evidence for DNA gyrase involvement in decatenation. Cell 36:1018–1088, 1984.

    Google Scholar 

  35. Sugino A, Peebles CL, Kreuzer KN, Cozzarelli NR: 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. Proc Natl Acad Sci USA 74:4761–4771, 1977.

    Google Scholar 

  36. Thompson RJ, Mosig G: An ATP-dependent supercoiling topoisomerase of Chlamydomonas reinhardii affects accumulation of specific chloroplast transcripts. Nucl Acids Res 13:873–891, 1985.

    Google Scholar 

  37. Tse Y, Wang JC: E. coli and M. luteus DNA topoisomerase I can catalyze catenation or decatenation of double-stranded DNA rings. Cell 22:269–276, 1980.

    Google Scholar 

  38. Wang JC, Liu LF: DNA topoisomerases: Enzymes that catalyze the concerted breaking and rejoining of DNA backbone bonds. Molecular Genetics, Part III. pp 65–88, Academic Press, New York, 1979.

    Google Scholar 

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  1. Biochemical Laboratory, Kobe Women's University, Suma-ku, 654, Kobe, Japan

    Hideki Fukata, Kazue Ohgami & Hirosuke Fukasawa

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  1. Hideki Fukata
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  2. Kazue Ohgami
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  3. Hirosuke Fukasawa
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Fukata, H., Ohgami, K. & Fukasawa, H. Isolation and characterization of DNA topoisomerase II from cauliflower inflorescences. Plant Mol Biol 6, 137–144 (1986). https://doi.org/10.1007/BF00021482

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  • DOI: https://doi.org/10.1007/BF00021482

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