Skip to main content

Advertisement

SpringerLink
Log in

Genome Instability in Ataxia Telangiectasia (A-T) Families: Camptothecin-Induced Damage to Replicating DNA Discriminates between Obligate A-T Heterozygotes, A-T Homozygotes and Controls

  • Published:
Bioscience Reports

Previously we used the topoisomerase I inhibitor camptothecin (CPT), which kills mainly S-phase cells primarily by inducing double strand breaks (DSBs) in replication forks, to show that ataxia telangiectasia (A-T) fibroblasts are defective in the repair of this particular subclass of DSBs. CPT treated A-T cells reaching G2 have abnormally high levels of chromatid exchanges, viewed as prematurely condensed G2 chromosomes (G2 PCC), compared with normal cells where aberrations are mostly chromatid breaks. Here we show that A-T lymphoblastoid cells established from individuals with different mutations in the ATM gene also exhibit increased levels of chromosomal exchanges in response to CPT, indicating that the replication-associated DSBs are misrepaired in all these cells. From family studies we show that the presence of a single mutated allele in obligate A-T heterozygotes leads to intermediate levels of chromosomal exchanges in CPT-treated lymphoblastoid cells, thus providing a functional and sensitive assay to identify these individuals.

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

Abbreviations

A-T:

ataxia telangiectasia

CPT:

camptothecin

DSB:

double strand break

IR:

ionizing radiation

PCC:

prematurely condensed chromosomes.

References

  1. M. F. Lavin Y. Shiloh (1997) ArticleTitleThe genetic defect in ataxia telangiectasia Annu. Rev. Immunol. 15 177–202 Occurrence Handle10.1146/annurev.immunol.15.1.177 Occurrence Handle9143686

    Article  PubMed  Google Scholar 

  2. J. Thacker (1994) ArticleTitleCellular radiosensitivity in ataxia telangiectasia. Int. J. Radiat. Biol. 66 587–596

    Google Scholar 

  3. J. Houldsworth M. F. Lavin (1980) ArticleTitleEffect of ionizing radiation on DNA synthesis in ataxia telangiectasia cells Nucleic Acids Res. 8 3709–3720 Occurrence Handle7433105

    PubMed  Google Scholar 

  4. R. B. Painter B. R. Young (1980) ArticleTitleRadiosensitivity in ataxia telangiectasia: a new explanation Proc. Natl. Acad. Sci. USA 77 7315–7317 Occurrence Handle6938978

    PubMed  Google Scholar 

  5. K. Savitsky A. Bar-Shira A. Gilad G. Rotman Y. Ziv L. Vanagaite D. A. Tagle S. Smith T. Uziel S. Sfez M. Ashkenazi I. Pecker M. Frydman R. Harnik S. R. Patanjali A. Simmons G. A. Clines A. Sartiel R. A. Gatti L. Chessa O. Sanal M. F. Lavin N. G. J. Jaspers A. Malcolm R. Taylor C. R. Arlett T. Miki S. M. Weissman M. Lovett F. S. Collins Y. Shiloh (1995) ArticleTitleA single ataxia telangiectasia gene with a product similar to PI-3 kinase Science 268 1749–1753 Occurrence Handle7792600

    PubMed  Google Scholar 

  6. Y. Shiloh (2003) ArticleTitleATM and related protein kinases:safeguarding genome integrity Natl. Rev. Cancer 3 155–168 Occurrence Handle10.1038/nrc1011

    Article  Google Scholar 

  7. R. T. Johnson E. Gotoh A. M. Mullinger A. J. Ryan Y. Shiloh Y. Ziv S. Squires (1999) ArticleTitleTargeting double strand breaks to replicating DNA identifies a subpathway of DSB repair that is defective in ataxia telangiectasia cells Biochem. Biophys. Res. Co. 261 317–325 Occurrence Handle10.1006/bbrc.1999.1024

    Article  Google Scholar 

  8. A. J. Ryan S. Squires H. L. Strutt R. T. Johnson (1991) ArticleTitleCamptothecin cytotoxicity in mammalian cells is associated with the induction of persistent double strand breaks in replicating DNA Nucleic Acids Res. 19 3295–3300 Occurrence Handle1905803

    PubMed  Google Scholar 

  9. P. J. Smith T. A. Makinson J. V. Watson (1989) ArticleTitleEnhanced sensitivity to camptothecin in ataxia telangiectasia cells and its relationship with the expression of DNA topisomerase I Int. J. Radiat. Biol. 55 217–231 Occurrence Handle2563396

    PubMed  Google Scholar 

  10. L. Chessa P. Petrinelli A. Antonelli M. Fiorilli R. Elli L. Marcucci A. Federico E. Gandini (1992) ArticleTitleHeterogeneity in ataxia telangiectasia: classical phenotype associated with intermediate cellular radiosensitivity Am. J. Med. Genet. 42 741–746 Occurrence Handle10.1002/ajmg.1320420524 Occurrence Handle1632451

    Article  PubMed  Google Scholar 

  11. S. Gilad T. Khosravi D. Shkedy T. Uziel Y. Ziv K. Savitsky G. Rotman S. Smith L. Chessa T. J. Jorgensen R. Harnik M. Frydman O. Sanal S. Portnoi Z. Goldwicz N. G. Jaspers R. A. Gatti G. Lenoir M. F. Lavin K. Tatsumi R. D. Wegner Y. Shiloh A. Bar-Shira (1996) ArticleTitlePredominance of null mutations in ataxia telangiectasia Hum. Mol. Genet. 5 433–439 Occurrence Handle10.1093/hmg/5.4.433 Occurrence Handle8845835

    Article  PubMed  Google Scholar 

  12. S. Gilad L. Chessa R. Khosravi P. Russell Y. Galanty M. Piane R. A. Gatti T. J. Jorgensen Y. Shiloh A. Bar-Shira (1998) ArticleTitleGenotype-phenotype relationships in ataxia telangiectasia and variants Am. J. Hum. Genet. 62 551–561 Occurrence Handle10.1086/301755 Occurrence Handle9497252

    Article  PubMed  Google Scholar 

  13. J. Wright S. Teraoka S. Onengut A. Tolun R. A. Gatti H. D. Ochs P. Concannon (1996) ArticleTitleA high frequency of distinct ATM gene mutations in ataxia telangiectasia Am. J. Hum. Genet. 59 839–846 Occurrence Handle8808599

    PubMed  Google Scholar 

  14. M. Telatar Z. Wang N. Udar T. Liang E. Bernatowska-Matuszkiewicz M. Lavin Y. Shiloh P. Concannon R. A. Good R. A. Gatti (1996) ArticleTitleAtaxia-telangiectasia mutations in ATM cDNA detected by protein-truncation screening Am. J. Hum. Genet. 59 40–44 Occurrence Handle8659541

    PubMed  Google Scholar 

  15. Mitelman, F. S. (ed.) (1995) ISCN 1995 An International System for Human Cytogenetic Nomenclature. Karger Publishers Inc., Farmington, CT

  16. T. Stankovic A. M. J. Kidd A. Sutcliffe G. M. McGuire P. Robinson P. Weber T. Bedenham A. R. Bradwell D. F. Easton G. G. Lennox N. Haites P. J. Byrd A. M. R. Taylor (1998) ArticleTitleATM mutations and phenotypes in ataxia telangiectasia families in the British Isles: expression of mutant ATM and the risk of leukemia, lymphoma, and breast cancer Am. J. Hum. Genet. 62 334–345 Occurrence Handle10.1086/301706 Occurrence Handle9463314

    Article  PubMed  Google Scholar 

  17. S. Squires A. J. Ryan H. L. Strutt R. T. Johnson (1993) ArticleTitleHypersensitivity of Cockayne’s Syndrome cells to camptothecin is associated with the generation of abnormally high levels of double strand breaks in nascent DNA Cancer Res. 53 2012–2019 Occurrence Handle7683249

    PubMed  Google Scholar 

  18. A. J. Ryan S. Squires H. Strutt A. Evans R. T. Johnson (1994) ArticleTitleDifferent fates of camptothecin-induced replication fork-associated double-strand DNA breaks in mammalian cells Carcinogenesis 15 823–828 Occurrence Handle8200082

    PubMed  Google Scholar 

  19. D. Morrell C. L. Chase M. Swift (1990) ArticleTitleCancers in 44 families with ataxia telangiectasia Cancer Genet. Cytogenet. 50 119–123 Occurrence Handle10.1016/0165-4608(90)90245-6 Occurrence Handle2253179

    Article  PubMed  Google Scholar 

  20. M. Swift D. Morrell R. B. Massey C. L. Chase (1991) ArticleTitleIncidence of cancer in 161 families affected by ataxia telangiectasia N. Engl. J. Med. 325 1831–1836 Occurrence Handle1961222

    PubMed  Google Scholar 

  21. M. Swift (2001) ArticleTitlePublic health burden of cancer in ataxia telangiectasia heterozygotes. J. Natl. Cancer I. 93 84–85 Occurrence Handle10.1093/jnci/93.2.84

    Article  Google Scholar 

  22. M. Lavin (1998) ArticleTitleRole of the ataxia telangiectasia gene (ATM) in breast cancer. Br. Med. J. 317 486–487

    Google Scholar 

  23. P. Athma R. Rappaport M. Swift (1996) ArticleTitleMolecular genotyping shows that ataxia telangiectasia heterozygotes are predisposed to breast cancer Cancer Genet. Cytogenet. 92 130–134 Occurrence Handle10.1016/S0165-4608(96)00328-7 Occurrence Handle8976369

    Article  PubMed  Google Scholar 

  24. H. M. Inskip L. J. Kinlen A. M. Taylor C. G. Woods C. F. Arlett (1999) ArticleTitleRisk of breast cancer and other cancers in heterozygotes for ataxia telangiectasia Br. J. Cancer 79 1304–1307 Occurrence Handle10.1038/sj.bjc.6690209 Occurrence Handle10098776

    Article  PubMed  Google Scholar 

  25. J. H. Olson (2001) ArticleTitleCancer patients with ataxia telangiectasia and their relatives in the Nordic countries. J. Natl. Cancer I 93 121–127 Occurrence Handle10.1093/jnci/93.2.121

    Article  Google Scholar 

  26. M. Swift P. J. Reitnauer D. Morrell C. L. Chase (1987) ArticleTitleBreast and other cancers in families with ataxia telangiectasia N. Engl. J. Med. 316 1289–1294 Occurrence Handle3574400

    PubMed  Google Scholar 

  27. K. Spring F. Ahangari S. P. Scott P. Waring D. M. Purdie P. C. Chen K. Hourigan J. Ramsay P. J. McKinnon M. Swift M. F. Lavin (2002) ArticleTitleMice heterozygous for mutation in Atm the gene involved in ataxia telangiectasia, have heightened susceptibility to cancer Nat. Genet. 32 185–190 Occurrence Handle10.1038/ng958 Occurrence Handle12195425

    Article  PubMed  Google Scholar 

  28. K. Heikkinen S.-M. Karppinen Y. Soini M. Makinen R. Winquist (2003) ArticleTitleMutation screening of Mre11 complex genes: indication of RAD50 involvement in breast and ovarian cancer susceptibility J. Med. Genet. 40 e131 Occurrence Handle10.1136/jmg.40.12.e131 Occurrence Handle14684699

    Article  PubMed  Google Scholar 

  29. A. Eastman (2004) ArticleTitleCell cycle checkpoints and their impact on anticancer therapeutic strategies. J. Cell. Biochem. 91 223–231 Occurrence Handle10.1002/jcb.10699 Occurrence Handle14743382

    Article  PubMed  Google Scholar 

  30. T. K. Pandita W. N. Hittelman (1994) ArticleTitleIncreased initial levels of chromosome damage and heterogeneous chromosome repair in ataxia telangiectasia heterozygote cells Mutat. Res. 310 1–13 Occurrence Handle7523872

    PubMed  Google Scholar 

  31. A. Tchirkov J.-O. Bay D. Perin Y.-J. Bignon P. Rio M. Grancho F. Kwiatkowski M. Giollant P. Malet P. Verrelle (1997) ArticleTitleDetection of heterozygous carriers of the ataxia telangiectasia (ATM) gene by G2 phase chromosomal radiosensitivity of peripheral blood lymphocytes Hum. Genet. 101 312–316 Occurrence Handle10.1007/s004390050634 Occurrence Handle9439660

    Article  PubMed  Google Scholar 

  32. C. S. Djuzenova D. Schindler H. Stopper H. Hoehn M. Flentje U. Oppitz (1999) ArticleTitleIdentification of ataxia telangiectasia heterozygotes, a cancer-prone population, using the single-cell gel electrophoresis (comet) assay Lab. Invest. 79 699–705 Occurrence Handle10378512

    PubMed  Google Scholar 

  33. M. Stumm S. Neubauer S. Keindorff R.-D. Wegner P. Wieacker R. Sauer (2001) ArticleTitleHigh frequency of spontaneous translocations revealed by FISH in cells from patients with the cancer-prone syndromes ataxia telangiectasia and Nijmegen breakage syndrome Cytogenet. Cell Genet. 92 186–191 Occurrence Handle10.1159/000056900 Occurrence Handle11435685

    Article  PubMed  Google Scholar 

  34. S. Neubauer R. Arutyunyan M. Stumm T. Dörk R. Bendix M. Bremer R. Varon R. Sauer E. Gebhart (2002) ArticleTitleRadiosensitivity of ataxia telangiectasia and Nijmegen breakage syndrome homozygotes and heterozygotes as determined by three-color FISH chromosome painting Radiat. Res. 157 312–321 Occurrence Handle11839094

    PubMed  Google Scholar 

  35. M. Fernet N. Moullan A. Lauge D. Stoppa-Lyonnet J. Hall (2004) ArticleTitleCellular responses to ionising radiation of AT heterozygotes; differences between missense and truncating mutation carriers Br. J. Cancer 90 866–873 Occurrence Handle10.1038/sj.bjc.6601549 Occurrence Handle14970866

    Article  PubMed  Google Scholar 

  36. S. Gutiérrez-Enriquez M. Fernet T. Dörk M. Bremer A. Lauge D. Stoppa- Lyonnet N. Moullan S. Angèle J. Hall (2004) ArticleTitleFunctional consequences of ATM sequence variants for chromosomal radiosensitivity Genes Chromosomes Cancer 40 109–119 Occurrence Handle10.1002/gcc.20025 Occurrence Handle15101044

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Coriell Cell Repositories, Coriell Institute for Medical Research, 403 Haddon Avenue, Camden, New Jersey, 08103, USA

    Jay C. Leonard, Ann M. Mullinger, John Schmidt & Robert T. Johnson

  2. Department of Zoology, University of Cambridge, Downing Street, Cambridge, CB2 3EJ, UK

    Ann M. Mullinger

  3. JDRF/WT Diabetes and Inflammation Laboratory, Department of Medical Genetics, University of Cambridge, Cambridge, CB2 2XY, UK

    Heather J. Cordell

  4. Mary Lyon Centre, Medical Research Council, Harwell, Didcot, Oxfordshire, OX11 ORD, UK

    Robert T. Johnson

Authors
  1. Jay C. Leonard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ann M. Mullinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Heather J. Cordell
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Robert T. Johnson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Robert T. Johnson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leonard, J.C., Mullinger, A.M., Schmidt, J. et al. Genome Instability in Ataxia Telangiectasia (A-T) Families: Camptothecin-Induced Damage to Replicating DNA Discriminates between Obligate A-T Heterozygotes, A-T Homozygotes and Controls. Biosci Rep 24, 617–629 (2004). https://doi.org/10.1007/s10540-005-2796-6

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10540-005-2796-6

Keywords

Search

Navigation