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. 1983;2(8):1303–1308. doi: 10.1002/j.1460-2075.1983.tb01585.x

Purification and characterization of Xenopus laevis topoisomerase II.

P Benedetti 1, M I Baldi 1, E Mattoccia 1, G P Tocchini-Valentini 1
PMCID: PMC555276  PMID: 10872324

Abstract

We have purified to apparent homogeneity a type II DNA topoisomerase from Xenopus laevis oocyte nuclei (germinal vesicles, or GV). The most pure preparations contain a single polypeptide of 175,000 daltons as determined by SDS-gel electrophoresis. The enzyme changes the linking number of DNA circles in steps of two and reversibly knots or catenates DNA rings. No gyrase activity is detectable and ATP is required.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Attardi D. G., De Paolis A., Tocchini-Valentini G. P. Purification and characterization of Xenopus laevis type I topoisomerase. J Biol Chem. 1981 Apr 25;256(8):3654–3661. [PubMed] [Google Scholar]
  2. Baase W. A., Wang J. C. An omega protein from Drosophila melanogaster. Biochemistry. 1974 Oct 8;13(21):4299–4303. doi: 10.1021/bi00718a009. [DOI] [PubMed] [Google Scholar]
  3. Baldi M. I., Benedetti P., Mattoccia E., Tocchini-Valentini G. P. In vitro catenation and decatenation of DNA and a novel eucaryotic ATP-dependent topoisomerase. Cell. 1980 Jun;20(2):461–467. doi: 10.1016/0092-8674(80)90632-7. [DOI] [PubMed] [Google Scholar]
  4. Brown P. O., Cozzarelli N. R. A sign inversion mechanism for enzymatic supercoiling of DNA. Science. 1979 Nov 30;206(4422):1081–1083. doi: 10.1126/science.227059. [DOI] [PubMed] [Google Scholar]
  5. Champoux J. J., Dulbecco R. An activity from mammalian cells that untwists superhelical DNA--a possible swivel for DNA replication (polyoma-ethidium bromide-mouse-embryo cells-dye binding assay). Proc Natl Acad Sci U S A. 1972 Jan;69(1):143–146. doi: 10.1073/pnas.69.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Champoux J. J., McConaughy B. L. Purification and characterization of the DNA untwisting enzyme from rat liver. Biochemistry. 1976 Oct 19;15(21):4638–4642. doi: 10.1021/bi00666a014. [DOI] [PubMed] [Google Scholar]
  7. Champoux J. J. Proteins that affect DNA conformation. Annu Rev Biochem. 1978;47:449–479. doi: 10.1146/annurev.bi.47.070178.002313. [DOI] [PubMed] [Google Scholar]
  8. Cozzarelli N. R. DNA gyrase and the supercoiling of DNA. Science. 1980 Feb 29;207(4434):953–960. doi: 10.1126/science.6243420. [DOI] [PubMed] [Google Scholar]
  9. Cozzarelli N. R. DNA topoisomerases. Cell. 1980 Nov;22(2 Pt 2):327–328. doi: 10.1016/0092-8674(80)90341-4. [DOI] [PubMed] [Google Scholar]
  10. Dynan W. S., Jendrisak J. J., Hager D. A., Burgess R. R. Purification and characterization of wheat germ DNA topoisomerase I (nicking-closing enzyme). J Biol Chem. 1981 Jun 10;256(11):5860–5865. [PubMed] [Google Scholar]
  11. Gellert M. DNA topoisomerases. Annu Rev Biochem. 1981;50:879–910. doi: 10.1146/annurev.bi.50.070181.004311. [DOI] [PubMed] [Google Scholar]
  12. Gellert M., Mizuuchi K., O'Dea M. H., Nash H. A. DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3872–3876. doi: 10.1073/pnas.73.11.3872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hsieh T., Brutlag D. ATP-dependent DNA topoisonmerase from D. melanogaster reversibly catenates duplex DNA rings. Cell. 1980 Aug;21(1):115–125. doi: 10.1016/0092-8674(80)90119-1. [DOI] [PubMed] [Google Scholar]
  14. Keller W. Characterization of purified DNA-relaxing enzyme from human tissue culture cells. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2550–2554. doi: 10.1073/pnas.72.7.2550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kreuzer K. N., Cozzarelli N. R. Formation and resolution of DNA catenanes by DNA gyrase. Cell. 1980 May;20(1):245–254. doi: 10.1016/0092-8674(80)90252-4. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Liu L. F., Liu C. C., Alberts B. M. Type II DNA topoisomerases: enzymes that can unknot a topologically knotted DNA molecule via a reversible double-strand break. Cell. 1980 Mar;19(3):697–707. doi: 10.1016/s0092-8674(80)80046-8. [DOI] [PubMed] [Google Scholar]
  18. Mattoccia E., Attardi D. G., Tocchini-Valentini G. P. DNA-relaxing activity and endonuclease activity in Xenopus laevis oocytes. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4551–4554. doi: 10.1073/pnas.73.12.4551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mattoccia E., Baldi M. I., Carrara G., Fruscoloni P., Benedetti P., Tocchini-Valentini G. P. Separation of RNA transcription and processing activities from X. laevis germinal vesicles. Cell. 1979 Nov;18(3):643–648. doi: 10.1016/0092-8674(79)90119-3. [DOI] [PubMed] [Google Scholar]
  20. Miller K. G., Liu L. F., Englund P. T. A homogeneous type II DNA topoisomerase from HeLa cell nuclei. J Biol Chem. 1981 Sep 10;256(17):9334–9339. [PubMed] [Google Scholar]
  21. Mizuuchi K., Fisher L. M., O'Dea M. H., Gellert M. DNA gyrase action involves the introduction of transient double-strand breaks into DNA. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1847–1851. doi: 10.1073/pnas.77.4.1847. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Oakley B. R., Kirsch D. R., Morris N. R. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. doi: 10.1016/0003-2697(80)90470-4. [DOI] [PubMed] [Google Scholar]
  23. Scalenghe F., Buscaglia M., Steinheil C., Crippa M. Large scale isolation of nuclei and nucleoli from vitellogenic oocytes of Xenopus laevis. Chromosoma. 1978 May 16;66(4):299–308. doi: 10.1007/BF00328531. [DOI] [PubMed] [Google Scholar]
  24. Staudenbauer W. L., Orr E. DNA gyrase: affinity chromatography on novobiocin-Sepharose and catalytic properties. Nucleic Acids Res. 1981 Aug 11;9(15):3589–3603. doi: 10.1093/nar/9.15.3589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sugino A., Higgins N. P., Brown P. O., Peebles C. L., Cozzarelli N. R. Energy coupling in DNA gyrase and the mechanism of action of novobiocin. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4838–4842. doi: 10.1073/pnas.75.10.4838. [DOI] [PMC free article] [PubMed] [Google Scholar]

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