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:

Chk2 is required for optimal mitotic delay in response to irradiation-induced DNA damage incurred in G2 phase

Oncogene volume 27, pages 896–906 (2008)Cite this article

Abstract

Whether Chk2 contributes to DNA damage-induced arrest in G2 has been controversial. To investigate this issue further, we generated Chk2-deficient DT40 B-lymphoma cells by gene targeting and compared their cell cycle response to ionizing radiation (IR) with wild-type (WT) and isogenic Chk1-deficient counterparts. After moderate doses of IR (4 Gy), we find that Chk2−/− cells which are in G1 or S phase at the time of irradiation arrest efficiently in G2. In contrast, Chk2−/− cells which are in G2 when DNA damage is incurred exhibit an impaired mitotic delay compared to WT, with the result that cells enter mitosis with damaged DNA as judged by the presence of numerous γ-H2AX foci on condensed chromosomes. Impaired G2 delay as the result of Chk2 deficiency can be detected at very low doses of radiation (0.1 Gy), and may allow division with spontaneous DNA damage, since a higher proportion of mitotic Chk2−/− cells bear spontaneous γ-H2AX foci and damaged chromosomes during unperturbed growth compared to WT. The contribution of Chk2 to G2/M delay is epistatic to that of Chk1, since Chk1−/− cells exhibit no measurable mitotic delay at any radiation dose tested. We suggest that this function of Chk2 could contribute to tumour suppression, since cell division with low levels of spontaneous damage is likely to promote genetic instability and thus carcinogenesis.

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
Figure 8

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  • Ahn JY, Schwarz JK, Piwnica-Worms H, Canman CE . (2000). Threonine 68 phosphorylation by ataxia telangiectasia mutated is required for efficient activation of Chk2 in response to ionizing radiation. Cancer Res 60: 5934–5936.

    CAS  PubMed  Google Scholar 

  • Bartek J, Lukas J . (2003). Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell 3: 421–429.

    Article  CAS  Google Scholar 

  • Bartkova J, Horejsi Z, Koed K, Kramer A, Tort F, Zieger K et al. (2005). DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature 434: 864–870.

    Article  CAS  Google Scholar 

  • Bell DW, Varley JM, Szydlo TE, Kang DH, Wahrer DC, Shannon KE et al. (1999). Heterozygous germ line hCHK2 mutations in Li–Fraumeni syndrome. Science 286: 2528–2531.

    Article  CAS  Google Scholar 

  • Cao L, Kim S, Xiao C, Wang RH, Coumoul X, Wang X et al. (2006). ATM-Chk2-p53 activation prevents tumorigenesis at an expense of organ homeostasis upon Brca1 deficiency. EMBO J 25: 2167–2177.

    Article  CAS  Google Scholar 

  • Chen Y, Sanchez Y . (2004). Chk1 in the DNA damage response: conserved roles from yeasts to mammals. DNA Repair (Amst) 3: 1025–1032.

    Article  CAS  Google Scholar 

  • Hirao A, Cheung A, Duncan G, Girard PM, Elia AJ, Wakeham A et al. (2002). Chk2 is a tumor suppressor that regulates apoptosis in both an ataxia telangiectasia mutated (ATM)-dependent and an ATM-independent manner. Mol Cell Biol 22: 6521–6532.

    Article  CAS  Google Scholar 

  • Hirao A, Kong YY, Matsuoka S, Wakeham A, Ruland J, Yoshida H et al. (2000). DNA damage-induced activation of p53 by the checkpoint kinase Chk2. Science 287: 1824–1827.

    Article  CAS  Google Scholar 

  • Jallepalli PV, Lengauer C, Vogelstein B, Bunz F . (2003). The Chk2 tumor suppressor is not required for p53 responses in human cancer cells. J Biol Chem 278: 20475–20479.

    Article  CAS  Google Scholar 

  • Jazayeri A, Falck J, Lukas C, Bartek J, Smith GC, Lukas J et al. (2006). ATM- and cell cycle-dependent regulation of ATR in response to DNA double-strand breaks. Nat Cell Biol 8: 37–45.

    Article  CAS  Google Scholar 

  • Lukas C, Bartkova J, Latella L, Falck J, Mailand N, Schroeder T et al. (2001). DNA damage-activated kinase Chk2 is independent of proliferation or differentiation yet correlates with tissue biology. Cancer Res 61: 4990–4993.

    CAS  Google Scholar 

  • Mailand N, Bekker-Jensen S, Bartek J, Lukas J . (2006). Destruction of Claspin by SCFbetaTrCP restrains Chk1 activation and facilitates recovery from genotoxic stress. Mol Cell 23: 307–318.

    Article  CAS  Google Scholar 

  • Matsuoka S, Huang M, Elledge SJ . (1998). Linkage of ATM to cell cycle regulation by the Chk2 protein kinase. Science 282: 1893–1897.

    Article  CAS  Google Scholar 

  • McGowan CH . (2002). Checking in on Cds1 (Chk2): a checkpoint kinase and tumor suppressor. Bioessays 24: 502–511.

    Article  CAS  Google Scholar 

  • Miller CW, Ikezoe T, Krug U, Hofmann WK, Tavor S, Vegesna V et al. (2002). Mutations of the CHK2 gene are found in some osteosarcomas, but are rare in breast, lung, and ovarian tumors. Genes Chromosomes Cancer 33: 17–21.

    Article  CAS  Google Scholar 

  • Nevanlinna H, Bartek J . (2006). The CHEK2 gene and inherited breast cancer susceptibility. Oncogene 25: 5912–5919.

    Article  CAS  Google Scholar 

  • O'Connell MJ, Walworth NC, Carr AM . (2000). The G2-phase DNA-damage checkpoint. Trends Cell Biol 10: 296–303.

    Article  CAS  Google Scholar 

  • Peschiaroli A, Dorrello NV, Guardavaccaro D, Venere M, Halazonetis T, Sherman NE et al. (2006). SCFbetaTrCP-mediated degradation of Claspin regulates recovery from the DNA replication checkpoint response. Mol Cell 23: 319–329.

    Article  CAS  Google Scholar 

  • Sancar A, Lindsey-Boltz LA, Unsal-Kacmaz K, Linn S . (2004). Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu Rev Biochem 73: 39–85.

    Article  CAS  Google Scholar 

  • Shiromizu T, Goto H, Tomono Y, Bartek J, Totsukawa G, Inoko A et al. (2006). Regulation of mitotic function of Chk1 through phosphorylation at novel sites by cyclin-dependent kinase 1 (Cdk1). Genes Cells 11: 477–485.

    Article  CAS  Google Scholar 

  • Skoufias DA, Lacroix FB, Andreassen PR, Wilson L, Margolis RL . (2004). Inhibition of DNA decatenation, but not DNA damage, arrests cells at metaphase. Mol Cell 15: 977–990.

    Article  CAS  Google Scholar 

  • Sonoda E, Sasaki MS, Buerstedde JM, Bezzubova O, Shinohara A, Ogawa H et al. (1998). Rad51-deficient vertebrate cells accumulate chromosomal breaks prior to cell death. EMBO J 17: 598–608.

    Article  CAS  Google Scholar 

  • Sorensen CS, Syljuasen RG, Falck J, Schroeder T, Ronnstrand L, Khanna KK et al. (2003). Chk1 regulates the S phase checkpoint by coupling the physiological turnover and ionizing radiation-induced accelerated proteolysis of Cdc25A. Cancer Cell 3: 247–258.

    Article  CAS  Google Scholar 

  • Takai H, Naka K, Okada Y, Watanabe M, Harada N, Saito S et al. (2002). Chk2-deficient mice exhibit radioresistance and defective p53-mediated transcription. EMBO J 21: 5195–5205.

    Article  CAS  Google Scholar 

  • Takao N, Kato H, Mori R, Morrison C, Sonada E, Sun X et al. (1999). Disruption of ATM in p53-null cells causes multiple functional abnormalities in cellular response to ionizing radiation. Oncogene 18: 7002–7009.

    Article  CAS  Google Scholar 

  • Xu B, Kim ST, Lim DS, Kastan MB . (2002). Two molecularly distinct G(2)/M checkpoints are induced by ionizing irradiation. Mol Cell Biol 22: 1049–1059.

    Article  CAS  Google Scholar 

  • Yu Q, Rose JH, Zhang H, Pommier Y . (2001). Antisense inhibition of Chk2/hCds1 expression attenuates DNA damage-induced S and G2 checkpoints and enhances apoptotic activity in HEK-293 cells. FEBS Lett 505: 7–12.

    Article  CAS  Google Scholar 

  • Zachos G, Rainey MD, Gillespie DA . (2003). Chk1-deficient tumour cells are viable but exhibit multiple checkpoint and survival defects. EMBO J 22: 713–723.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Gareth Inman for insightful comments on the manuscript. This work was supported by Cancer Research UK (CR-UK) and the Association for International Cancer Research (AICR).

Author information

Author notes

  1. M D Rainey

    Present address: 3Present address: Department of Hematology–Oncology, St Jude Children's Research Hospital, Memphis, TN 38105, USA.,

Authors and Affiliations

  1. Beatson Institute for Cancer Research, Garscube Estate, Glasgow, UK

    M D Rainey, E J Black, G Zachos & D A F Gillespie

  2. Faculty of Biomedical and Life Sciences, University of Glasgow, Glasgow, UK

    D A F Gillespie

Authors
  1. M D Rainey
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E J Black
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G Zachos
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D A F Gillespie
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D A F Gillespie.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rainey, M., Black, E., Zachos, G. et al. Chk2 is required for optimal mitotic delay in response to irradiation-induced DNA damage incurred in G2 phase. Oncogene 27, 896–906 (2008). https://doi.org/10.1038/sj.onc.1210702

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210702

Keywords

This article is cited by

Search

Advanced search

Quick links