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:

Chk1-dependent slowing of S-phase progression protects DT40 B-lymphoma cells against killing by the nucleoside analogue 5-fluorouracil

Oncogene volume 25, pages 5359–5369 (2006)Cite this article

Abstract

Chk1 plays a crucial role in the DNA damage and replication checkpoints in vertebrates and may therefore be an important determinant of tumour cell responses to genotoxic anticancer drugs. To evaluate this concept we compared the effects of the nucleoside analogue 5-fluorouracil (5FU) on cell cycle progression and clonogenic survival in DT40 B-lymphoma cells with an isogenic mutant derivative in which Chk1 function was ablated by gene targeting. We show that 5FU activates Chk1 in wild-type DT40 cells and that 5FU-treated cells accumulate in the S phase of the cell cycle due to slowing of the overall rate of DNA replication. In marked contrast, Chk1-deficient DT40 cells fail to slow DNA replication upon initial exposure to 5FU, despite equivalent inhibition of the target enzyme thymidylate synthase, and instead accumulate progressively in the G1 phase of the following cell cycle. This G1 accumulation cannot be reversed rapidly by exogenous thymidine or removal of 5FU, and is associated with increased incorporation of 5FU into genomic DNA and severely diminished clonogenic survival. Taken together, these results demonstrate that a Chk1-dependent replication checkpoint which slows S phase progression can protect tumour cells against the cytotoxic effects of 5FU.

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

References

  • Abraham RT . (2001). Genes Dev 15: 2177–2196.

  • Bartek J, Lukas J . (2003). Cancer Cell 3: 421–429.

  • Feijoo C, Hall-Jackson C, Wu R, Jenkins D, Leitch J, Gilbert DM et al. (2001). J Cell Biol 154: 913–923.

  • Glazer RI, Lloyd LS . (1982). Mol Pharmacol 21: 468–473.

  • Graves PR, Yu L, Schwarz JK, Gales J, Sausville EA, O'Connor PM et al. (2000). J Biol Chem 275: 5600–5605.

  • Heidelberger C, Danenberg PV, Moran RG . (1983). Adv Enzymol Relat Areas Mol Biol 54: 58–119.

  • Ingraham HA, Dickey L, Goulian M . (1986). Biochemistry 25: 3225–3230.

  • Ingraham HA, Tseng BY, Goulian M . (1980). Cancer Res 40: 998–1001.

  • Kastan MB, Bartek J . (2004). Nature 432: 316–323.

  • Kufe DW, Major PP . (1981). J Biol Chem 256: 9802–9805.

  • Liu X, Guo Y, Li Y, Jiang Y, Chubb S, Azuma A et al (2005). Cancer Res 65: 6874–6881.

  • Longley DB, Harkin DP, Johnston PG . (2003). Nat Rev Cancer 3: 330–338.

  • Mauro DJ, De Riel JK, Tallarida RJ, Sirover MA . (1993). Mol Pharmacol 43: 854–857.

  • Meyers M, Wagner MW, Hwang HS, Kinsella TJ, Boothman DA . (2001). Cancer Res 61: 5193–5201.

  • Meyers M, Wagner MW, Mazurek A, Schmutte C, Fishel R, Boothman DA . (2005). J Biol Chem 280: 5516–5526.

  • Morgan MA, Parsels LA, Parsels JD, Mesiwala AK, Maybaum J, Lawrence TS . (2005). Cancer Res 65: 6835–6842.

  • Nyberg KA, Michelson RJ, Putnam CW, Weinert TA . (2002). Annu Rev Genet 36: 617–656.

  • Rhind N, Russell P . (2000). J Cell Sci 113 (Part 22): 3889–3896.

  • Sampath D, Rao VA, Plunkett W . (2003). Oncogene 22: 9063–9074.

  • Santi DV, McHenry CS, Sommer H . (1974). Biochemistry 13: 471–481.

  • Shi Z, Azuma A, Sampath D, Li YX, Huang P, Plunkett W . (2001). Cancer Res 61: 1065–1072.

  • Tajima A, Hess MT, Cabrera BL, Kolodner RD, Carethers JM . (2004). Gastroenterology 127: 1678–1684.

  • Xiao Z, Xue J, Sowin TJ, Rosenberg SH, Zhang H . (2005). Oncogene 24: 1403–1411.

  • Zachos G, Rainey MD, Gillespie DA . (2003). EMBO J 22: 713–723.

  • Zachos G, Rainey MD, Gillespie DA . (2005). Mol Cell Biol 25: 563–574.

  • Zhao H, Piwnica-Worms H . (2001). Mol Cell Biol 21: 4129–4139.

  • Zou L, Elledge SJ . (2003). Science 300: 1542–1548.

Download references

Acknowledgements

We thank Dr Kevin Ryan 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. R Jones

    Present address: School of Medical Sciences, University Walk, Clifton, Bristol, BS8 1TD, UK

  2. H M R Robinson and R Jones: These authors contributed equally to this work.

Authors and Affiliations

  1. Beatson Institute for Cancer Research, Glasgow, UK

    H M R Robinson, M Walker, G Zachos & D A F Gillespie

  2. CRUK Centre for Oncology and Applied Pharmacology, Beatson Laboratories, University of Glasgow, Glasgow, UK

    H M R Robinson, R Jones, R Brown & J Cassidy

  3. Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, UK

    D A F Gillespie

Authors
  1. H M R Robinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R Jones
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Walker
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G Zachos
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R Brown
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J Cassidy
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. 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

Robinson, H., Jones, R., Walker, M. et al. Chk1-dependent slowing of S-phase progression protects DT40 B-lymphoma cells against killing by the nucleoside analogue 5-fluorouracil. Oncogene 25, 5359–5369 (2006). https://doi.org/10.1038/sj.onc.1209532

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1209532

Keywords

This article is cited by

Search

Advanced search

Quick links