Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Nov 11;20(21):5587–5592. doi: 10.1093/nar/20.21.5587

Isolation of cDNA clones encoding the beta isozyme of human DNA topoisomerase II and localisation of the gene to chromosome 3p24.

J R Jenkins 1, P Ayton 1, T Jones 1, S L Davies 1, D L Simmons 1, A L Harris 1, D Sheer 1, I D Hickson 1
PMCID: PMC334390  PMID: 1333583

Abstract

Topoisomerases catalyse the interconversion of topological isomers of DNA and have key roles in nucleic acid metabolism. Human cells express two distinct type II topoisomerase isozymes, designated topoisomerase II alpha (170 kDa form) and topoisomerase II beta (180 kDa form). We have isolated cDNA clones encoding the beta isozyme from a human B-cell library. The proposed coding region for the topoisomerase II beta protein is 4,863 nucleotides long and would encode a polypeptide with a calculated M(r) of 182,705. The predicted topoisomerase II beta protein sequence shows striking similarity (72% identical residues) to that of the human alpha isozyme, and homology to topoisomerase II proteins from Drosophila, yeast and bacteria. Regions of greatest amino acid sequence divergence lie at the extreme N-terminus and over a C-terminal domain comprising approximately 25% of the total protein. We have quantified the level of topoisomerase II beta mRNA in a panel of human tumour cell lines of different origin using an RNase protection assay, and compared the level to that of topoisomerase II alpha mRNA. Topoisomerase II beta mRNA was expressed in haemopoietic, epithelial and fibroblast cell lines, although to different extents, with U937 cells (promonocytic leukaemia) showing a particularly high level. There was no obvious relationship in terms of level of expression between the topoisomerase II alpha and beta genes. We have localised the gene encoding topoisomerase II beta protein to chromosome 3p24 in the human genome.

Full text

PDF
5587

Images in this article

Selected References

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

  1. Adachi Y., Luke M., Laemmli U. K. Chromosome assembly in vitro: topoisomerase II is required for condensation. Cell. 1991 Jan 11;64(1):137–148. doi: 10.1016/0092-8674(91)90215-k. [DOI] [PubMed] [Google Scholar]
  2. Austin C. A., Fisher L. M. Isolation and characterization of a human cDNA clone encoding a novel DNA topoisomerase II homologue from HeLa cells. FEBS Lett. 1990 Jun 18;266(1-2):115–117. doi: 10.1016/0014-5793(90)81520-x. [DOI] [PubMed] [Google Scholar]
  3. Bae Y. S., Kawasaki I., Ikeda H., Liu L. F. Illegitimate recombination mediated by calf thymus DNA topoisomerase II in vitro. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2076–2080. doi: 10.1073/pnas.85.7.2076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brill S. J., Sternglanz R. Transcription-dependent DNA supercoiling in yeast DNA topoisomerase mutants. Cell. 1988 Jul 29;54(3):403–411. doi: 10.1016/0092-8674(88)90203-6. [DOI] [PubMed] [Google Scholar]
  5. Cardenas M. E., Dang Q., Glover C. V., Gasser S. M. Casein kinase II phosphorylates the eukaryote-specific C-terminal domain of topoisomerase II in vivo. EMBO J. 1992 May;11(5):1785–1796. doi: 10.1002/j.1460-2075.1992.tb05230.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Christman M. F., Dietrich F. S., Fink G. R. Mitotic recombination in the rDNA of S. cerevisiae is suppressed by the combined action of DNA topoisomerases I and II. Cell. 1988 Nov 4;55(3):413–425. doi: 10.1016/0092-8674(88)90027-x. [DOI] [PubMed] [Google Scholar]
  7. Chung T. D., Drake F. H., Tan K. B., Per S. R., Crooke S. T., Mirabelli C. K. Characterization and immunological identification of cDNA clones encoding two human DNA topoisomerase II isozymes. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9431–9435. doi: 10.1073/pnas.86.23.9431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cook P. R. The nucleoskeleton and the topology of replication. Cell. 1991 Aug 23;66(4):627–635. doi: 10.1016/0092-8674(91)90109-c. [DOI] [PubMed] [Google Scholar]
  9. Davies S. M., Robson C. N., Davies S. L., Hickson I. D. Nuclear topoisomerase II levels correlate with the sensitivity of mammalian cells to intercalating agents and epipodophyllotoxins. J Biol Chem. 1988 Nov 25;263(33):17724–17729. [PubMed] [Google Scholar]
  10. 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 U S A. 1984 May;81(9):2616–2620. doi: 10.1073/pnas.81.9.2616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dillehay L. E., Jacobson-Kram D., Williams J. R. DNA topoisomerases and models of sister-chromatid exchange. Mutat Res. 1989 Nov;215(1):15–23. doi: 10.1016/0027-5107(89)90213-3. [DOI] [PubMed] [Google Scholar]
  12. Drake F. H., Hofmann G. A., Bartus H. F., Mattern M. R., Crooke S. T., Mirabelli C. K. Biochemical and pharmacological properties of p170 and p180 forms of topoisomerase II. Biochemistry. 1989 Oct 3;28(20):8154–8160. doi: 10.1021/bi00446a029. [DOI] [PubMed] [Google Scholar]
  13. Drake F. H., Zimmerman J. P., McCabe F. L., Bartus H. F., Per S. R., Sullivan D. M., Ross W. E., Mattern M. R., Johnson R. K., Crooke S. T. Purification of topoisomerase II from amsacrine-resistant P388 leukemia cells. Evidence for two forms of the enzyme. J Biol Chem. 1987 Dec 5;262(34):16739–16747. [PubMed] [Google Scholar]
  14. 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]
  15. Fernandes D. J., Danks M. K., Beck W. T. Decreased nuclear matrix DNA topoisomerase II in human leukemia cells resistant to VM-26 and m-AMSA. Biochemistry. 1990 May 1;29(17):4235–4241. doi: 10.1021/bi00469a028. [DOI] [PubMed] [Google Scholar]
  16. Friche E., Danks M. K., Schmidt C. A., Beck W. T. Decreased DNA topoisomerase II in daunorubicin-resistant Ehrlich ascites tumor cells. Cancer Res. 1991 Aug 15;51(16):4213–4218. [PubMed] [Google Scholar]
  17. Gasser S. M., Laroche T., Falquet J., Boy de la Tour E., Laemmli U. K. Metaphase chromosome structure. Involvement of topoisomerase II. J Mol Biol. 1986 Apr 20;188(4):613–629. doi: 10.1016/s0022-2836(86)80010-9. [DOI] [PubMed] [Google Scholar]
  18. Giaccone G., Gazdar A. F., Beck H., Zunino F., Capranico G. Multidrug sensitivity phenotype of human lung cancer cells associated with topoisomerase II expression. Cancer Res. 1992 Apr 1;52(7):1666–1674. [PubMed] [Google Scholar]
  19. Glikin G. C., Blangy D. In vitro transcription by Xenopus oocytes RNA polymerase III requires a DNA topoisomerase II activity. EMBO J. 1986 Jan;5(1):151–155. doi: 10.1002/j.1460-2075.1986.tb04189.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Goto T., Wang J. C. Cloning of yeast TOP1, the gene encoding DNA topoisomerase I, and construction of mutants defective in both DNA topoisomerase I and DNA topoisomerase II. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7178–7182. doi: 10.1073/pnas.82.21.7178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hinds M., Deisseroth K., Mayes J., Altschuler E., Jansen R., Ledley F. D., Zwelling L. A. Identification of a point mutation in the topoisomerase II gene from a human leukemia cell line containing an amsacrine-resistant form of topoisomerase II. Cancer Res. 1991 Sep 1;51(17):4729–4731. [PubMed] [Google Scholar]
  22. Hirano T., Konoha G., Toda T., Yanagida M. Essential roles of the RNA polymerase I largest subunit and DNA topoisomerases in the formation of fission yeast nucleolus. J Cell Biol. 1989 Feb;108(2):243–253. doi: 10.1083/jcb.108.2.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Holm C., Goto T., Wang J. C., Botstein D. DNA topoisomerase II is required at the time of mitosis in yeast. Cell. 1985 Jun;41(2):553–563. doi: 10.1016/s0092-8674(85)80028-3. [DOI] [PubMed] [Google Scholar]
  24. Kemp B. E., Pearson R. B. Protein kinase recognition sequence motifs. Trends Biochem Sci. 1990 Sep;15(9):342–346. doi: 10.1016/0968-0004(90)90073-k. [DOI] [PubMed] [Google Scholar]
  25. Kim R. A., Wang J. C. A subthreshold level of DNA topoisomerases leads to the excision of yeast rDNA as extrachromosomal rings. Cell. 1989 Jun 16;57(6):975–985. doi: 10.1016/0092-8674(89)90336-x. [DOI] [PubMed] [Google Scholar]
  26. Klein F., Laroche T., Cardenas M. E., Hofmann J. F., Schweizer D., Gasser S. M. Localization of RAP1 and topoisomerase II in nuclei and meiotic chromosomes of yeast. J Cell Biol. 1992 Jun;117(5):935–948. doi: 10.1083/jcb.117.5.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Negri C., Chiesa R., Cerino A., Bestagno M., Sala C., Zini N., Maraldi N. M., Astaldi Ricotti G. C. Monoclonal antibodies to human DNA topoisomerase I and the two isoforms of DNA topoisomerase II: 170- and 180-kDa isozymes. Exp Cell Res. 1992 Jun;200(2):452–459. doi: 10.1016/0014-4827(92)90195-e. [DOI] [PubMed] [Google Scholar]
  30. Osheroff N., Zechiedrich E. L., Gale K. C. Catalytic function of DNA topoisomerase II. Bioessays. 1991 Jun;13(6):269–273. doi: 10.1002/bies.950130603. [DOI] [PubMed] [Google Scholar]
  31. Pommier Y., Schwartz R. E., Zwelling L. A., Kohn K. W. Effects of DNA intercalating agents on topoisomerase II induced DNA strand cleavage in isolated mammalian cell nuclei. Biochemistry. 1985 Nov 5;24(23):6406–6410. doi: 10.1021/bi00344a014. [DOI] [PubMed] [Google Scholar]
  32. Snapka R. M. Topoisomerase inhibitors can selectively interfere with different stages of simian virus 40 DNA replication. Mol Cell Biol. 1986 Dec;6(12):4221–4227. doi: 10.1128/mcb.6.12.4221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sperry A. O., Blasquez V. C., Garrard W. T. Dysfunction of chromosomal loop attachment sites: illegitimate recombination linked to matrix association regions and topoisomerase II. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5497–5501. doi: 10.1073/pnas.86.14.5497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tan K. B., Dorman T. E., Falls K. M., Chung T. D., Mirabelli C. K., Crooke S. T., Mao J. Topoisomerase II alpha and topoisomerase II beta genes: characterization and mapping to human chromosomes 17 and 3, respectively. Cancer Res. 1992 Jan 1;52(1):231–234. [PubMed] [Google Scholar]
  35. Tsai-Pflugfelder M., Liu L. F., Liu A. A., Tewey K. M., Whang-Peng J., Knutsen T., Huebner K., Croce C. M., Wang J. C. Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7177–7181. doi: 10.1073/pnas.85.19.7177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Uemura T., Ohkura H., Adachi Y., Morino K., Shiozaki K., Yanagida M. DNA topoisomerase II is required for condensation and separation of mitotic chromosomes in S. pombe. Cell. 1987 Sep 11;50(6):917–925. doi: 10.1016/0092-8674(87)90518-6. [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]
  38. Webb C. D., Latham M. D., Lock R. B., Sullivan D. M. Attenuated topoisomerase II content directly correlates with a low level of drug resistance in a Chinese hamster ovary cell line. Cancer Res. 1991 Dec 15;51(24):6543–6549. [PubMed] [Google Scholar]
  39. Williams S. V., Jones T. A., Cottrell S., Zehetner G., Varesco L., Ward T., Thomas H., Lawson P. A., Solomon E., Bodmer W. F. Fine mapping of probes in the adenomatous polyposis coli region of chromosome 5 by in situ hybridization. Genes Chromosomes Cancer. 1991 Sep;3(5):382–389. doi: 10.1002/gcc.2870030509. [DOI] [PubMed] [Google Scholar]
  40. Woessner R. D., Mattern M. R., Mirabelli C. K., Johnson R. K., Drake F. H. Proliferation- and cell cycle-dependent differences in expression of the 170 kilodalton and 180 kilodalton forms of topoisomerase II in NIH-3T3 cells. Cell Growth Differ. 1991 Apr;2(4):209–214. [PubMed] [Google Scholar]
  41. Wyckoff E., Natalie D., Nolan J. M., Lee M., Hsieh T. Structure of the Drosophila DNA topoisomerase II gene. Nucleotide sequence and homology among topoisomerases II. J Mol Biol. 1989 Jan 5;205(1):1–13. doi: 10.1016/0022-2836(89)90361-6. [DOI] [PubMed] [Google Scholar]
  42. Yang L., Wold M. S., Li J. J., Kelly T. J., Liu L. F. Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro. Proc Natl Acad Sci U S A. 1987 Feb;84(4):950–954. doi: 10.1073/pnas.84.4.950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zini N., Martelli A. M., Sabatelli P., Santi S., Negri C., Astaldi Ricotti G. C., Maraldi N. M. The 180-kDa isoform of topoisomerase II is localized in the nucleolus and belongs to the structural elements of the nucleolar remnant. Exp Cell Res. 1992 Jun;200(2):460–466. doi: 10.1016/0014-4827(92)90196-f. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES