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. 1986 Oct;83(19):7152–7156. doi: 10.1073/pnas.83.19.7152

ATP-independent type II topoisomerase from trypanosomes.

S Douc-Rasy, A Kayser, J F Riou, G Riou
PMCID: PMC386673  PMID: 3020537

Abstract

We have characterized in Trypanosoma cruzi a DNA topoisomerase capable of decatenating complex trypanosomal kinetoplast DNA networks in the absence of ATP. The enzymatic activity requires Mg2+ and K+. Using a defined DNA topoisomer we showed that the linking number changes by steps of 2, which characterizes the enzyme as a type II topoisomerase. The enzyme can catenate supercoiled DNA molecules, unknot DNA, and cleave double-stranded DNA. The enzyme has no ATPase activity. The native enzyme has an Mr of about 200,000. Crude extracts and partially purified fractions contain an aggregating factor that can substitute spermidine in catenating reactions. Because of the presence of this factor, the kinetoplast DNA can only be decatenated by purified fractions. The enzyme is inhibited by certain drugs and provides a potential target for chemotherapy. Such an enzyme was also characterized in Trypanosoma equiperdum.

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Selected References

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

  1. Andrews P. Estimation of molecular size and molecular weights of biological compounds by gel filtration. Methods Biochem Anal. 1970;18:1–53. [PubMed] [Google Scholar]
  2. Arai K., Yasuda S., Kornberg A. Mechanism of dnaB protein action. I. Crystallization and properties of dnaB protein, an essential replication protein in Escherichia coli. J Biol Chem. 1981 May 25;256(10):5247–5252. [PubMed] [Google Scholar]
  3. Cockerill P. N., Garrard W. T. Chromosomal loop anchorage of the kappa immunoglobulin gene occurs next to the enhancer in a region containing topoisomerase II sites. Cell. 1986 Jan 31;44(2):273–282. doi: 10.1016/0092-8674(86)90761-0. [DOI] [PubMed] [Google Scholar]
  4. Douc-Rasy S., Kayser A., Riou G. F. Inhibition of the reactions catalysed by a type I topoisomerase and catenating enzyme of Trypanosoma cruzi by DNA-intercalating drugs. Preferential inhibition of the catenating reaction. EMBO J. 1984 Jan;3(1):11–16. doi: 10.1002/j.1460-2075.1984.tb01754.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Douc-Rasy S., Kayser A., Riou G. A specific inhibitor of type I DNA-topoisomerase of Trypanosoma cruzi: dimethyl-hydroxy-ellipticinium. Biochem Biophys Res Commun. 1983 Nov 30;117(1):1–5. doi: 10.1016/0006-291x(83)91532-2. [DOI] [PubMed] [Google Scholar]
  6. Douc-Rasy S., Multon E., Kayser A., Riou G. Inhibition par les dérivés de l'ellipticine des réactions catalysées par les topoisomérases: inhibition préférentielle d'une topoisomérase II. C R Seances Acad Sci III. 1983;296(19):899–904. [PubMed] [Google Scholar]
  7. Douc-Rasy S., Riou J. F., Kayser A., Riou G. Une ADN-topoisomérase de trypanosome capable de caténer, décaténer et dénouer l'ADN sans ATP. C R Acad Sci III. 1986;302(8):283–286. [PubMed] [Google Scholar]
  8. Drlica K. Biology of bacterial deoxyribonucleic acid topoisomerases. Microbiol Rev. 1984 Dec;48(4):273–289. doi: 10.1128/mr.48.4.273-289.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Duguet M., Lavenot C., Harper F., Mirambeau G., De Recondo A. M. DNA topoisomerases from rat liver: physiological variations. Nucleic Acids Res. 1983 Feb 25;11(4):1059–1075. doi: 10.1093/nar/11.4.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Earnshaw W. C., Halligan B., Cooke C. A., Heck M. M., Liu L. F. Topoisomerase II is a structural component of mitotic chromosome scaffolds. J Cell Biol. 1985 May;100(5):1706–1715. doi: 10.1083/jcb.100.5.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Englund P. T., Hajduk S. L., Marini J. C. The molecular biology of trypanosomes. Annu Rev Biochem. 1982;51:695–726. doi: 10.1146/annurev.bi.51.070182.003403. [DOI] [PubMed] [Google Scholar]
  12. Forterre P., Mirambeau G., Jaxel C., Nadal M., Duguet M. High positive supercoiling in vitro catalyzed by an ATP and polyethylene glycol-stimulated topoisomerase from Sulfolobus acidocaldarius. EMBO J. 1985 Aug;4(8):2123–2128. doi: 10.1002/j.1460-2075.1985.tb03902.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gellert M. DNA topoisomerases. Annu Rev Biochem. 1981;50:879–910. doi: 10.1146/annurev.bi.50.070181.004311. [DOI] [PubMed] [Google Scholar]
  14. Gellert M., Mizuuchi K., O'Dea M. H., Itoh T., Tomizawa J. I. Nalidixic acid resistance: a second genetic character involved in DNA gyrase activity. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4772–4776. doi: 10.1073/pnas.74.11.4772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Glikin G. C., Ruberti I., Worcel A. Chromatin assembly in Xenopus oocytes: in vitro studies. Cell. 1984 May;37(1):33–41. doi: 10.1016/0092-8674(84)90298-8. [DOI] [PubMed] [Google Scholar]
  16. Goto T., Wang J. C. Yeast DNA topoisomerase II is encoded by a single-copy, essential gene. Cell. 1984 Apr;36(4):1073–1080. doi: 10.1016/0092-8674(84)90057-6. [DOI] [PubMed] [Google Scholar]
  17. Goto T., Wang J. C. 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. 1982 May 25;257(10):5866–5872. [PubMed] [Google Scholar]
  18. 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]
  19. Krasnow M. A., Cozzarelli N. R. Catenation of DNA rings by topoisomerases. Mechanism of control by spermidine. J Biol Chem. 1982 Mar 10;257(5):2687–2693. [PubMed] [Google Scholar]
  20. Liu L. F., Davis J. L., Calendar R. Novel topologically knotted DNA from bacteriophage P4 capsids: studies with DNA topoisomerases. Nucleic Acids Res. 1981 Aug 25;9(16):3979–3989. doi: 10.1093/nar/9.16.3979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liu L. F., Rowe T. C., Yang L., Tewey K. M., Chen G. L. Cleavage of DNA by mammalian DNA topoisomerase II. J Biol Chem. 1983 Dec 25;258(24):15365–15370. [PubMed] [Google Scholar]
  22. 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]
  23. Nelson E. M., Tewey K. M., Liu L. F. Mechanism of antitumor drug action: poisoning of mammalian DNA topoisomerase II on DNA by 4'-(9-acridinylamino)-methanesulfon-m-anisidide. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1361–1365. doi: 10.1073/pnas.81.5.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Osheroff N., Shelton E. R., Brutlag D. L. DNA topoisomerase II from Drosophila melanogaster. Relaxation of supercoiled DNA. J Biol Chem. 1983 Aug 10;258(15):9536–9543. [PubMed] [Google Scholar]
  25. Riou G. F., 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. 1985 Feb 1;146(3):483–488. doi: 10.1111/j.1432-1033.1985.tb08677.x. [DOI] [PubMed] [Google Scholar]
  26. Riou G. F., Gabillot M., Douc-Rasy S., Kayser A., Barrois M. A type I DNA topoisomerase from Trypanosoma cruzi. Eur J Biochem. 1983 Aug 15;134(3):479–484. doi: 10.1111/j.1432-1033.1983.tb07592.x. [DOI] [PubMed] [Google Scholar]
  27. Riou G. F., Yot P. Heterogeneity of the kinetoplast DNA molecules of Trypanosoma cruzi. Biochemistry. 1977 May 31;16(11):2390–2396. doi: 10.1021/bi00630a013. [DOI] [PubMed] [Google Scholar]
  28. Riou J. F., Gabillot M., Philippe M., Schrevel J., Riou G. Purification and characterization of Plasmodium berghei DNA topoisomerases I and II: drug action, inhibition of decatenation and relaxation, and stimulation of DNA cleavage. Biochemistry. 1986 Apr 8;25(7):1471–1479. doi: 10.1021/bi00355a001. [DOI] [PubMed] [Google Scholar]
  29. Sander M., Hsieh T. Double strand DNA cleavage by type II DNA topoisomerase from Drosophila melanogaster. J Biol Chem. 1983 Jul 10;258(13):8421–8428. [PubMed] [Google Scholar]
  30. Saucier J. M., Benard J., da Silva J., Riou G. Occurrence of a kinetoplast DNA-protein complex in Trypanosoma cruzi. Biochem Biophys Res Commun. 1981 Aug 14;101(3):988–994. doi: 10.1016/0006-291x(81)91846-5. [DOI] [PubMed] [Google Scholar]
  31. Shlomai J., Zadok A. Reversible decatenation of kinetoplast DNA by a DNA topoisomerase from trypanosomatids. Nucleic Acids Res. 1983 Jun 25;11(12):4019–4034. doi: 10.1093/nar/11.12.4019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shore D., Langowski J., Baldwin R. L. DNA flexibility studied by covalent closure of short fragments into circles. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4833–4837. doi: 10.1073/pnas.78.8.4833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Siegel L. M., Monty K. J. Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim Biophys Acta. 1966 Feb 7;112(2):346–362. doi: 10.1016/0926-6585(66)90333-5. [DOI] [PubMed] [Google Scholar]
  34. Sugino A., Peebles C. L., Kreuzer K. N., Cozzarelli N. R. 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 U S A. 1977 Nov;74(11):4767–4771. doi: 10.1073/pnas.74.11.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tse Y. C., Kirkegaard K., Wang J. C. Covalent bonds between protein and DNA. Formation of phosphotyrosine linkage between certain DNA topoisomerases and DNA. J Biol Chem. 1980 Jun 25;255(12):5560–5565. [PubMed] [Google Scholar]
  36. Udvardy A., Schedl P., Sander M., Hsieh T. S. Novel partitioning of DNA cleavage sites for Drosophila topoisomerase II. Cell. 1985 Apr;40(4):933–941. doi: 10.1016/0092-8674(85)90353-8. [DOI] [PubMed] [Google Scholar]
  37. Wang J. C. DNA topoisomerases. Annu Rev Biochem. 1985;54:665–697. doi: 10.1146/annurev.bi.54.070185.003313. [DOI] [PubMed] [Google Scholar]

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