Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Topoisomerase IIα maintains genomic stability through decatenation G2 checkpoint signaling

Oncogene volume 29, pages 4787–4799 (2010)Cite this article

Subjects

Abstract

Topoisomerase IIα (topoIIα) is an essential mammalian enzyme that topologically modifies DNA and is required for chromosome segregation during mitosis. Previous research suggests that inhibition of topoII decatenatory activity triggers a G2 checkpoint response, which delays mitotic entry because of insufficient decatenation of daughter chromatids. Here we examine the effects of both topoIIα and topoIIβ on decatenatory activity in cell extracts, DNA damage and decatenation G2 checkpoint function, and the frequencies of p16INK4A allele loss and gain. In diploid human fibroblast lines, depletion of topoIIα by small-interfering RNA was associated with severely reduced decatenatory activity, delayed progression from G2 into mitosis and insensitivity to G2 arrest induced by the topoII catalytic inhibitor ICRF-193. Furthermore, interphase nuclei of topoIIα-depleted cells showed increased frequencies of losses and gains of the tumor suppressor genetic locus p16INK4A. This study shows that the topoIIα protein is required for decatenation G2 checkpoint function, and inactivation of decatenation and the decatenation G2 checkpoint leads to abnormal chromosome segregation and genomic instability.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

Abbreviations

topoII:

topoisomerase II

DSB:

double-strand break

NHDF:

normal human diploid fibroblast

NTC:

non-targeting control

MER:

mitotic entry rate

References

  • Akimitsu N, Adachi N, Hirai H, Hossain MS, Hamamoto H, Kobayashi M et al. (2003). Enforced cytokinesis without complete nuclear division in embryonic cells depleting the activity of DNA topoisomerase IIalpha. Genes Cells 8: 393–402.

    Article  CAS  PubMed  Google Scholar 

  • Austin CA, Marsh KL . (1998). Eukaryotic DNA topoisomerase II beta. Bioessays 20: 215–226.

    Article  CAS  PubMed  Google Scholar 

  • Bandele OJ, Osheroff N . (2007). Bioflavanoids as poisons of human topoisomerase IIα and IIβ. Biochemistry 46: 6097–6108.

    Article  CAS  PubMed  Google Scholar 

  • Bandele OJ, Osheroff N . (2008). (-)-epigallocatechin gallate, a major constituent of green tea, poisons human type II topoisomerases. Chem Res Toxicol 21: 936–943.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Barjesteh van Waalwijk van Doorn-Khosrovani S, Janssen J, Maas LM, Godschalk RWL, Nijhuis JG, van Schooten FJ . (2007). Dietary flavonoids induce MLL translocations in primary human CD34+ cells. Carcinogenesis 28: 1703–1709.

    Article  PubMed  Google Scholar 

  • Baxter J, Diffley JFX . (2008). Topoisomerase II inactivation prevents the completion of DNA replication in budding yeast. Mol Cell 30: 790–802.

    Article  CAS  PubMed  Google Scholar 

  • Beser AR, Tuzlali S, Guzey D, Dolec Guler S, Hacihanefioglu S, Dalay N . (2007). Her-2, TOP2A, and chromosome 17 alterations in breast cancer. Pathol Oncol Res 13: 180–185.

    Article  PubMed  Google Scholar 

  • Biersack H, Jensen S, Gromova I, Nielsen IS, Westergaard O, Andersen AH . (1996). Active heterodimers are formed from human DNA topoisomerase IIα and IIβ isoforms. Proc Natl Acad Sci USA 93: 8288–8293.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bower JJ, Zhou Y, Zhou T, Simpson DA, Arlander SJ, Paules RS et al. (2010). Revised genetic requirements for the decatenation G2 checkpoint: the role of ATM. Cell Cycle 9: 1617–1628.

    Article  CAS  PubMed  Google Scholar 

  • Burden DA, Osheroff N . (1998). Mechanism of action of eukaryotic topoisomerase II and drugs targeted to the enzyme. Biochim Biophys Acta 1400: 139–154.

    Article  CAS  PubMed  Google Scholar 

  • Capranico G, Tinelli S, Austin CA, Fisher ML, Zunino F . (1992). Different patterns of gene expression of topoisomerase II isoforms in differentiated tissues during murine development. Biochim Biophys Acta 1132: 43–48.

    Article  CAS  PubMed  Google Scholar 

  • Carpenter AJ, Porter AC . (2004). Construction, characterization, and complementation of a conditional-lethal DNA topoisomerase II alpha mutant human cell line. Mol Biol Cell 15: 5700–5711.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Coelho PA, Queiroz-Machado J, Carmo AM, Moutinho-Pereira S, Maiato H, Sunkel CE . (2008). Dual role of topoisomerase II in centromere resolution and aurora B activity. PLoS Biol 6: 1758–1777.

    Article  CAS  Google Scholar 

  • Cortes F, Pastor N, Mateos S, Dominguez I . (2003). Roles of DNA topoisomerases in chromosome segregation and mitosis. Mutat Res 543: 59–66.

    Article  CAS  PubMed  Google Scholar 

  • Damelin M, Sun YE, Sodja VB, Bestor TH . (2005). Decatenation checkpoint deficiency in stem and progenitor cells. Cancer Cell 8: 479–484.

    Article  CAS  PubMed  Google Scholar 

  • Dawlaty MM, Malureanu L, Jeganathan KB, Kao E, Sustmann C, Tahk S et al. (2008). Resolution of sister centromeres requires RanBP2-mediated SUMOylation of topoisomerase IIα. Cell 133: 103–115.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Deming PB, Cistulli CA, Zhao H, Graves PR, Piwnica-Worms H, Paules RS et al. (2001). The human decatenation checkpoint. Proc Natl Acad Sci USA 98: 12044–12049.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Deming PB, Flores KG, Downes CS, Paules RS, Kaufmann WK . (2002). ATR enforces the topoisomerase II-dependent G2 checkpoint through inhibition of Plk1 kinase. J Biol Chem 277: 36832–36838.

    Article  CAS  PubMed  Google Scholar 

  • Diaz-Martinez LA, Gimenez-Abian JF, Azuma Y, Guacci V, Gimenez-Martin G, Lanier LM et al. (2006). PIASgamma is required for faithful chromosome segregation in human cells. PLoS ONE 1: e53.

    Article  PubMed  PubMed Central  Google Scholar 

  • Doherty SC, McKeown SR, McKelvey-Martin V, Downes CS, Atala A, Yoo JJ et al. (2003). Cell cycle checkpoint function in bladder cancer. J Natl Cancer Inst 95: 1859–1868.

    Article  CAS  PubMed  Google Scholar 

  • Downes CS, Clarke DJ, Mullinger AM, Gimenez-Abian JF, Creighton AM, Johnson RT . (1994). A topoisomerase II-dependent G2 cycle checkpoint in mammalian cells (published erratum appears in Nature 1994; 372: 710). Nature 372: 467–470.

    Article  CAS  PubMed  Google Scholar 

  • Emmons M, Boulware D, Sullivan DM, Hazlehurst LA . (2006). Topoisomerase II beta levels are a determinant of melphalan-induced DNA crosslinks and sensitivity to cell death. Biochem Pharmacol 72: 11–18.

    Article  CAS  PubMed  Google Scholar 

  • Franchitto A, Oshima J, Pichierri P . (2003). The G2-phase decatenation checkpoint is defective in Werner syndrome cells. Cancer Res 63: 3289–3295.

    CAS  PubMed  Google Scholar 

  • Goswami PC, Roti Roti JL, Hunt CR . (1996). The cell cycle-coupled expression of topoisomerase IIα during S phase is regulated by mRNA stability and is disrupted by heat shock or ionizing radiation. Mol Cell Biol 16: 1500–1508.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gromova I, Biersack H, Jensen S, Nielsen OF, Westergaard O, Andersen AH . (1998). Characterization of DNA topoisomerase II α/β heterodimers in HeLa cells. Biochemistry 37: 16645–16652.

    Article  CAS  PubMed  Google Scholar 

  • Heck MMS, Hittelman WN, Earnshaw WC . (1988). Differential expression of DNA topoisomerases I and II during the eukaryotic cell cycle. Proc Natl Acad Sci USA 85: 1086–1090.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Holland AJ, Cleveland DW . (2009). Boveri revisited: chromosomal instability, aneuploidy, and tumorigenesis. Nat Rev Mol Cell Biol 10: 478–487.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jarvinen TAH, Kononen J, Pelto-Huikko M, Isola J . (1996). Expression of topoisomerase IIα is associated with rapid cell proliferation, aneuploidy, and c-erbB2 overexpression in breast cancer. Am J Pathol 148: 2073–2082.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Jensen LH, Nitiss KC, Rose A, Dong J, Zhou J, Hu T et al. (2000). A novel mechanism of cell killing by anti-topoisomerase II bisdioxopiperazines. J Biol Chem 275: 2137–2146.

    Article  CAS  PubMed  Google Scholar 

  • Johnson M, Phua HH, Bennett SC, Spence JM, Farr CJ . (2009). Studying vertebrate topoisomerase 2 function using a conditional knockdown system in DT40 cells. Nuc Acid Res 37: e98.

    Article  Google Scholar 

  • Ju BG, Lunyak VV, Perissi V, Garcia-Bassets I, Rose DW, Glass CK et al. (2006). A topoisomerase IIβ-mediated dsDNA break required for regulated transcription. Science 312: 1798–1802.

    Article  CAS  PubMed  Google Scholar 

  • Kaufmann WK, Schwartz JL, Hurt JC, Byrd LL, Galloway DA, Levedakou E et al. (1997). Inactivation of G2 checkpoint function and chromosomal destabilization are linked in human fibroblasts expressing human papillomavirus type 16 E6. Cell Growth Differ 8: 1105–1114.

    CAS  PubMed  Google Scholar 

  • Larsen AK, Skladanowski A, Bojanowski K . (1996). The roles of DNA topoisomerase II during the cell cycle. Prog Cell Cycle Res 2: 229–239.

    Article  CAS  PubMed  Google Scholar 

  • Li H, Wang Y, Liu X . (2008). Plk1-dependent phosphorylation regulates functions of DNA topoisomerase II in cell cycle progression. J Biol Chem 283: 6209–6221.

    Article  CAS  PubMed  Google Scholar 

  • Lorusso V, Manzione L, Silvestris N . (2007). Role of liposomal anthracyclines in breast cancer. Ann Oncol 18: vi70–vi73.

    PubMed  Google Scholar 

  • Lou Z, Minter-Dykhouse K, Chen J . (2005). BRCA1 participates in DNA decatenation. Nat Struct Mol Biol 12: 589–593.

    Article  CAS  PubMed  Google Scholar 

  • Luo K, Yuan J, Chen J, Lou Z . (2009). Topoisomerase IIα controls the decatenation checkpoint. Nat Cell Biol 11: 204–210.

    Article  CAS  PubMed  Google Scholar 

  • Lyu YL, Kerrigan JE, Lin CP, Azarova AM, Tsai YC, Ban Y et al. (2007). Topoisomerase IIβ mediated DNA double-strand breaks: implications in Doxorubicin Cardiotoxicity and prevention by Dexrazoxane. Cancer Res 67: 8839–8846.

    Article  CAS  PubMed  Google Scholar 

  • Maeshima K, Eltsov M, Laemmli UK . (2005). Chromosome structure: improved immunolabeling for electron microscopy. Chromosoma 114: 365–375.

    Article  PubMed  Google Scholar 

  • Mao Y, Desai SD, Ting CY, Hwang J, Liu LF . (2001). 26 S proteasome-mediated degradation of topoisomerase II cleavable complexes. J Biol Chem 276: 40652–40658.

    Article  CAS  PubMed  Google Scholar 

  • Nakagawa T, Hayashita Y, Maeno K, Masuda A, Sugito N, Osada H et al. (2004). Identification of decatenation G2 checkpoint impairment independently of DNA damage G2 checkpoint in human lung cancer cell lines. Cancer Res 64: 4826–4832.

    Article  CAS  PubMed  Google Scholar 

  • Park I, Avraham HK . (2006). Cell cycle-dependent DNA damage signaling induced by ICRF-193 involves ATM, ATR, CHK2, and BRCA1. Exp Cell Res 312: 1996–2008.

    Article  CAS  PubMed  Google Scholar 

  • Robinson HM, Bratlie-Thoresen S, Brown R, Gillespie DA . (2007). Chk1 is required for G2/M checkpoint response induced by the catalytic topoisomerase II inhibitor ICRF-193. Cell Cycle 6: 1265–1267.

    Article  CAS  PubMed  Google Scholar 

  • Roca J, Ishida R, Berger JM, Andoh T, Wang JC . (1994). Antitumor bisdioxopiperazines inhibit yeast DNA topoisomerase II by trapping the enzyme in the form of a closed protein clamp. Proc Natl Acad Sci USA 91: 1781–1785.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Santamaria A, Neef R, Eberspacher U, Eis K, Husemann M, Mumberg D et al. (2007). Use of the novel Plk1 inhibitor ZK-Thiazolidinone to elucidate functions of Plk1 in early and late stages of mitosis. Mol Biol Cell 18: 4024–4036.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sehested M, Jensen PB . (1996). Mapping of DNA topoisomerase II poisons (etoposide, clerocidin) and catalytic inhibitors (aclarubicin, ICRF-189) to four distinct steps in the topoisomerase II catalytic cycle. Biochem Pharm 51: 879–886.

    Article  CAS  PubMed  Google Scholar 

  • Simpson DA, Livanos E, Heffernan TP, Kaufmann WK . (2005). Telomerase exression is sufficient for chromosomal integrity in cells lacking p53-dependent G1 checkpoint function. J Carcinog 4: 18.

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang JC . (2002). Cellular roles of DNA topoisomerases: a molecular perspective. Nat Rev Mol Cell Biol 3: 430–440.

    Article  CAS  PubMed  Google Scholar 

  • Wang LHC, Schwarzbraun T, Speicher MR, Nigg EA . (2008). Persistence of DNA threads in human anaphase cells suggests late completion of sister chromatid decatenation. Chromosoma 117: 123–135.

    Article  PubMed  Google Scholar 

  • Xiao H, Mao Y, Desai SD, Zhou N, Ting CY, Hwang J et al. (2003). The topoisomerase II beta circular clamp arrests transcription and signals a 26S proteasome pathway. Proc Natl Acad Sci USA 100: 3239–3244.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou T, Chou JW, Simpson DA, Zhou Y, Mullen TE, Medeiro M et al. (2006). Profiles of global gene expression in ionizing-radiation-damaged human diploid fibroblasts reveal synchronization behind the G1 checkpoint in a G0-like state of quiescence. Environ Health Persp 114: 553–559.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Financial Support: This work was supported by an NIH-NCI Ruth L. Kirschstein National Research Service Award Grant # 1F32CA134155-01 (JJ Bower) and PHS grants CA81343 and P30-ES10126 (WK Kaufmann).

Author information

Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    J J Bower, G F Karaca, Y Zhou, D A Simpson, M Cordeiro-Stone & W K Kaufmann

  2. Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    J J Bower, M Cordeiro-Stone & W K Kaufmann

  3. Center for Environmental Health and Susceptibility, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    M Cordeiro-Stone & W K Kaufmann

Authors
  1. J J Bower
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G F Karaca
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D A Simpson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M Cordeiro-Stone
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. W K Kaufmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to W K Kaufmann.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bower, J., Karaca, G., Zhou, Y. et al. Topoisomerase IIα maintains genomic stability through decatenation G2 checkpoint signaling. Oncogene 29, 4787–4799 (2010). https://doi.org/10.1038/onc.2010.232

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2010.232

Keywords

This article is cited by

Search

Advanced search

Quick links