Skip to main content
SpringerLink
Log in

Organization of DNA topoisomerase II isotypes during the cell cycle of human lymphocytes and HeLa cells

  • Published:
Chromosome Research Aims and scope Submit manuscript

Abstract

We have monitored the organization of DNA topoisomerase II (Topo II) in relation to chromatin disaggregation during mitogen stimulation of lymphocytes and to the mitotic chromosome condensation cycle by immunofluorescence microscopy with isozyme-specific antibodies. Labelling for both Topo IIα and Topo IIβ was diffusely nucleoplasmic and non-nucleolar in resting lymphocytes and the pattern changed little during stimulation. Topo IIα labelling intensity increased in parallel with the extent of cell stimulation, but a fraction of fully stimulated cells was labelled very brightly. Topo IIβ labelling intensity was also greater in stimulated cells, but all partially and fully stimulated cells were labelled at the same, higher, intensity. In addition, anti-Topo IIβ detected a few small spots within nucleoli of stimulated cells that coincided with regions containing fibrillarin. In lymphocytes and HeLa, chromosome association of Topo IIα began in prophase and lasted throughout mitosis. In contrast, Topo IIβ stayed nucleoplasmic in prophase, was diffusely cytoplasmic during mitosis, and was first detected post-mitotically in nuclel with decondensing chromosomes and a reformed nuclear envelope. The results are consistent with a role for Topo IIα, but not for Topo IIβ, in mitotic chromosome condensation, and indicate that the isotypes may play independent roles in the reorganization of chromatin structure during lymphocyte mitogenic activation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Azum-Gélade M-C, Noaillac-Depeyre J, Caizerques-Ferrer M, Gas N (1994) Cell cycle redistribution of U3 snRNA and fibrillarin. Presence in the cytoplasmic nucleolus remnant and in the prenucleolar bodies at telophase.J Cell Sci 107: 463–475.

    PubMed  Google Scholar 

  • Berrios M, Osheroff N, Fisher PA (1985)In situ localization of DNA topoisomerase II, a major polypeptide component of theDrosophila nuclear matrix fraction.Proc Natl Acad Sci USA 82: 4142–4146.

    PubMed  Google Scholar 

  • Brown DL, Dudani R, Little JE et al. (1990) Dynamics of the nucleus during lymphocyte activation. In: E. Wang, J.-L. Wang, S. Chien, C.-W. Wu, eds.Biochemical and Structural Dynamics of the Cell Nucleus. New York: Academic Press, pp 161–170.

    Google Scholar 

  • Chaly N, Brown DL (1996) Is DNA topoisomerase IIβ a nucleolar protein?J Cell Biochem (in press).

  • Chaly N, Chen X (1993) Assembly of adenovirus-specific nuclear inclusions in lytically infected HeLa cells: an ultrastructural and cytochemical study.Biochem Cell Biol 71: 475–487.

    PubMed  Google Scholar 

  • Chaly N, Bladon T, Setterfield G et al. (1984) Redistribution of nuclear matrix antigens during the mitotic cell cycle.J Cell Biol 99: 661–671.

    Article  PubMed  Google Scholar 

  • Chaly N, Cadrin M, Kaplan JG, Brown DL (1988) Modulation of lymphocyte nuclear matrix organizationin vivo by 5, 6-dichloro-1-β-d-ribofuranosyl benzimidazole: an autoradiographic and immunofluorescence study.Biol Cell 63: 9–17.

    Article  PubMed  Google Scholar 

  • Chung TDY, Drake FH, Tan KB et al. (1989) Characterization and immunological identification of cDNA clones encoding two human DNA topoisomerase II isozymes.Proc Natl Acad Sci USA 86: 9431–9435.

    PubMed  Google Scholar 

  • Daev E, Chaly N, Brown DL et al. (1994) Role of topoisomerase II in the structural and functional evolution of mitogen-stimulated lymphocyte nuclei.Exp Cell Res 214: 331–342.

    Article  PubMed  Google Scholar 

  • Drake FH, Hofmann GA, Bartus HF et al. (1989) Biochemical and pharmacological properties of p170 and p180 forms of topoisomerase II.Biochemistry 28: 8154–8160.

    Article  PubMed  Google Scholar 

  • Earnshaw WC, Heck MMS (1985) Localization of Topoisomerase II in mitotic chromosomes.J Cell Biol 100: 1716–1725.

    Article  PubMed  Google Scholar 

  • Earnshaw WC, Halligan B, Cooke CA, Heck MMS, Liu LF (1985) Topoisomerase II is a structural component of mitotic chromosome scaffolds.J Cell Biol 100: 1716–1725.

    Article  PubMed  Google Scholar 

  • Gasser SM, Laroche T, Falquet J, Boy de la Tour E, Laemmli UK (1986) Metaphase chromosome structure: involvement of topoisomerase II.J Mol Biol 188: 613–629.

    Article  PubMed  Google Scholar 

  • Govoni M, Neri S, Labella T et al. (1995) Topoisomerase-II-mediated DNA cleavage within the human ribosomal genes.Biochem Biophys Res Commun 213: 282–288.

    Article  PubMed  Google Scholar 

  • Heck MMS, Earnshaw WC (1986) Topoisomerase II: a specific marker for cell proliferation.J Cell Biol 103: 2569–2581.

    Article  PubMed  Google Scholar 

  • Hirano T, Mitchison TJ (1993) Topoisomerase II does not play a scaffolding role in the organization of mitotic chromosomes assembled inXenopus egg extracts.J Cell Biol 120: 601–612.

    Article  PubMed  Google Scholar 

  • Kaplan JG, Brown DL, Chaly N et al. (1987) Structural and evolutionary implications of the packaging of DNA for differentiation and proliferation in the lymphocyte.J Mol Evol 26: 173–179.

    Article  PubMed  Google Scholar 

  • Kimura K, Nozaki N, Saijo M et al. (1994) Identification of the nature of modification that causes the shift of DNA Topoisomerase IIβ to apparent higher molecular weight forms in the M. Phase.J Biol Chem 269: 24523–24526.

    PubMed  Google Scholar 

  • Negri C, Chiesa R, Cerino A et al. (1992) Monoclonal antibodies to human DNA topoisomerase I and the two isoforms of DNA topoisomerase II: 170- and 180-kDa isozymes.Exp Cell Res 200: 452–459.

    Article  PubMed  Google Scholar 

  • Negri C, Scovassi AI, Braghetti A, Guano F, Astaldi-Ricotti GCB (1993) DNA topoisomerase IIβ: stability and distribution in different animal cells in comparison to DNA topoisomerase I and IIα.Exp Cell Res 206: 128–133.

    Article  PubMed  Google Scholar 

  • Ochs RL, Lischwe MA, Spohn WH, Busch H (1985) Fibrillarin: a new protein of the nucleolus identified by autoimmune sera.Biol Cell 54: 123–134.

    PubMed  Google Scholar 

  • Petrov P, Drake FH, Loranger A, Huang W, Hancock R (1993) Localization of DNA topoisomerase II in Chinese hamster fibroblasts by confocal and electron microscopy.Exp Cell Res 204: 73–81.

    Article  PubMed  Google Scholar 

  • Poljak L, Käs E (1995) Resolving the role of topoisomerase II in chromatin structure and function.Trends Cell Biol 5: 348–354.

    Article  PubMed  Google Scholar 

  • Prosperi E, Negri C, Marchese G, Astaldi-Ricotti GCB (1994) Expression of the 170-kDa and 180-kDa isoforms of DNA topoisomerase II in resting and proliferating human lymphocytes.Cell Prolif 27: 257–267.

    PubMed  Google Scholar 

  • Roy C, Brown DL, Little JE et al. (1992) The topoisomerase II inhibitor teniposide (VM-26) induces apoptosis in unstimulated mature murine lymphocytes.Exp Cell Res 200: 416–424.

    Article  PubMed  Google Scholar 

  • Saitoh N, Goldberg I, Earnshaw WC (1995) The SMC proteins and the coming of age of the chromosome scaffold hypothesis.BioEssays 17: 759–766.

    Article  PubMed  Google Scholar 

  • Scheer U, Weisenberger D (1994) The nucleolus.Curr Opin Cell Biol 6: 354–359.

    Article  PubMed  Google Scholar 

  • Setterfield G, Hall R, Bladon T, Little J, Kaplan JG (1983) Changes in structure and composition of lymphocyte nuclei during mitogenic stimulation.J Ultrastruct Res 82: 264–282.

    Article  PubMed  Google Scholar 

  • Setterfield G, Bladon T, Hall R et al. (1985) Extrachromatin nuclear components and structural changes in nuclei. In:Nuclear Envelope Structure and RNA Maturation. UCLA Symp. Mol. Cell. Biol. New Series Vol. 26. New York: Alan R. Liss, pp. 63–86.

    Google Scholar 

  • Sumner AT (1996) The distribution of topoisomerase II on mammalian chromosomes.Chrom Res 4: 5–14.

    Article  PubMed  Google Scholar 

  • Swedlow JR, Sedat JW, Agard DA (1993) Multiple chromosomal populations of Topoisomerase II detectedin vivo by time-lapse, three-dimensional wide-field microscopy.Cell 73: 97–108.

    Article  PubMed  Google Scholar 

  • Taagepera S, Rao PN, Drake FH, Gorbsky GJ (1993) DNA topoisomerase IIα is the major chromosome protein recognized by the mitotic phosphoprotein antibody MPM-2.Proc Natl Acad Sci USA 90: 8407–8411.

    PubMed  Google Scholar 

  • Tan KB, Dorman TE, Falls KM et al. (1992) Topoisomerase IIα and topoisomerase IIβ genes: characterization and mapping to human chromosomes 17 and 3, respectively.Cancer Res 52: 231–234.

    PubMed  Google Scholar 

  • Tsai-Pflugfelder M, Liu LF, Liu AA et al. (1988) Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22.Proc Natl Acad Sci USA 85: 7177–7181.

    PubMed  Google Scholar 

  • Watt PM, Hickson ID (1994) Structure and function of type II DNA topoisomerases.Biochem J 303: 681–695.

    PubMed  Google Scholar 

  • Woessner RD, Mattern MR, Mirabelli CK, Johnson RK, Drake FH (1991) Proliferation-cell cycle-dependent differences in expression of the 170kD and 180kD forms of topoisomerase II in NIH-3T3 cells.Cell Growth Differ 2: 209–214.

    PubMed  Google Scholar 

  • Zini N, Martelli AM, Sabatelli P et al. (1992) 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 204: 460–466.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, ELBA, Carleton University, 1125 Colonel By Drive, KIS 5B6, Ottawa, ON, Canada

    N. Chaly & X. Chen

  2. Department of Biology, University of Ottawa, Ottawa, ON, Canada

    J. Dentry & D. L. Brown

Authors
  1. N. Chaly
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. X. Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. Dentry
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. L. Brown
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

accepted for publication by T. D. Allen

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chaly, N., Chen, X., Dentry, J. et al. Organization of DNA topoisomerase II isotypes during the cell cycle of human lymphocytes and HeLa cells. Chromosome Res 4, 457–466 (1996). https://doi.org/10.1007/BF02265053

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02265053

Key words

Search

Navigation