Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1988 May 11;16(9):3919–3929. doi: 10.1093/nar/16.9.3919

Reduced DNA topoisomerase II activity in ataxia-telangiectasia cells.

S P Singh 1, R Mohamed 1, C Salmond 1, M F Lavin 1
PMCID: PMC336565  PMID: 2836804

Abstract

Considerable evidence supports a defect at the level of chromatin structure or recognition of that structure in cells from patients with the human genetic disorder ataxia-telangiectasia. Accordingly, we have investigated the activities of enzymes that alter the topology of DNA in Epstein Barr Virus-transformed lymphoblastoid cells from patients with this syndrome. Reduced activity of DNA topoisomerase II, determined by unknotting of P4 phage DNA, was observed in partially purified extracts from 5 ataxia-telangiectasia cell lines. The levels of enzyme activity was reduced substantially in 4 of these cell lines and to a lesser extent in the other cell line compared to controls. DNA topoisomerase I, assayed by relaxation of supercoiled DNA, was found to be present at comparable levels in both cell types. Reduced activity of topoisomerase II in ataxia-telangiectasia is compatible with the molecular, cellular and clinical changes described in this syndrome.

Full text

PDF
3919

Images in this article

3922Image
on p.3922
3924Image
on p.3924
3925Image
on p.3925
3926Image
on p.3926

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aurias A., Dutrillaux B., Buriot D., Lejeune J. High frequencies of inversions and translocations of chromosomes 7 and 14 in ataxia telangiectasia. Mutat Res. 1980 Feb;69(2):369–374. doi: 10.1016/0027-5107(80)90101-3. [DOI] [PubMed] [Google Scholar]
  2. Berrios M., Osheroff N., Fisher P. A. In situ localization of DNA topoisomerase II, a major polypeptide component of the Drosophila nuclear matrix fraction. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4142–4146. doi: 10.1073/pnas.82.12.4142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Chen P. C., Lavin M. F., Kidson C., Moss D. Identification of ataxia telangiectasia heterozygotes, a cancer prone population. Nature. 1978 Aug 3;274(5670):484–486. doi: 10.1038/274484a0. [DOI] [PubMed] [Google Scholar]
  5. Cockerill P. N., Garrard W. T. Chromosomal loop anchorage of the kappa immunoglobulin gene occurs next to the enhancer in a region containing topoisomerase II sites. Cell. 1986 Jan 31;44(2):273–282. doi: 10.1016/0092-8674(86)90761-0. [DOI] [PubMed] [Google Scholar]
  6. Cohen M. M., Shaham M., Dagan J., Shmueli E., Kohn G. Cytogenetic investigations in families with ataxia-telangiectasia. Cytogenet Cell Genet. 1975;15(5):338–356. doi: 10.1159/000130530. [DOI] [PubMed] [Google Scholar]
  7. Cornforth M. N., Bedford J. S. On the nature of a defect in cells from individuals with ataxia-telangiectasia. Science. 1985 Mar 29;227(4694):1589–1591. doi: 10.1126/science.3975628. [DOI] [PubMed] [Google Scholar]
  8. Cox R., Debenham P. G., Masson W. K., Webb M. B. Ataxia-telangiectasia: a human mutation giving high-frequency misrepair of DNA double-stranded scissions. Mol Biol Med. 1986 Jun;3(3):229–244. [PubMed] [Google Scholar]
  9. Cox R., Masson W. K., Debenham P. G., Webb M. B. The use of recombinant DNA plasmids for the determination of DNA-repair and recombination in cultured mammalian cells. Br J Cancer Suppl. 1984;6:67–72. [PMC free article] [PubMed] [Google Scholar]
  10. Debenham P. G., Webb M. B., Jones N. J., Cox R. Molecular studies on the nature of the repair defect in ataxia-telangiectasia and their implications for cellular radiobiology. J Cell Sci Suppl. 1987;6:177–189. doi: 10.1242/jcs.1984.supplement_6.12. [DOI] [PubMed] [Google Scholar]
  11. DiNardo S., Voelkel K., Sternglanz R. DNA topoisomerase II mutant of Saccharomyces cerevisiae: topoisomerase II is required for segregation of daughter molecules at the termination of DNA replication. Proc Natl Acad Sci U S A. 1984 May;81(9):2616–2620. doi: 10.1073/pnas.81.9.2616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Duguet M., Lavenot C., Harper F., Mirambeau G., De Recondo A. M. DNA topoisomerases from rat liver: physiological variations. Nucleic Acids Res. 1983 Feb 25;11(4):1059–1075. doi: 10.1093/nar/11.4.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Earnshaw W. C., Heck M. M. Localization of topoisomerase II in mitotic chromosomes. J Cell Biol. 1985 May;100(5):1716–1725. doi: 10.1083/jcb.100.5.1716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ford M. D., Martin L., Lavin M. F. The effects of ionizing radiation on cell cycle progression in ataxia telangiectasia. Mutat Res. 1984 Jan;125(1):115–122. doi: 10.1016/0027-5107(84)90038-1. [DOI] [PubMed] [Google Scholar]
  15. Halligan B. D., Edwards K. A., Liu L. F. Purification and characterization of a type II DNA topoisomerase from bovine calf thymus. J Biol Chem. 1985 Feb 25;260(4):2475–2482. [PubMed] [Google Scholar]
  16. Henner W. D., Blazka M. E. Hypersensitivity of cultured ataxia-telangiectasia cells to etoposide. J Natl Cancer Inst. 1986 Jun;76(6):1007–1011. [PubMed] [Google Scholar]
  17. Holm C., Goto T., Wang J. C., Botstein D. DNA topoisomerase II is required at the time of mitosis in yeast. Cell. 1985 Jun;41(2):553–563. doi: 10.1016/s0092-8674(85)80028-3. [DOI] [PubMed] [Google Scholar]
  18. Houldsworth J., Lavin M. F. Effect of ionizing radiation on DNA synthesis in ataxia telangiectasia cells. Nucleic Acids Res. 1980 Aug 25;8(16):3709–3720. doi: 10.1093/nar/8.16.3709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Liu L. F., Davis J. L., Calendar R. Novel topologically knotted DNA from bacteriophage P4 capsids: studies with DNA topoisomerases. Nucleic Acids Res. 1981 Aug 25;9(16):3979–3989. doi: 10.1093/nar/9.16.3979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Liu L. F., Rowe T. C., Yang L., Tewey K. M., Chen G. L. Cleavage of DNA by mammalian DNA topoisomerase II. J Biol Chem. 1983 Dec 25;258(24):15365–15370. [PubMed] [Google Scholar]
  21. McFarlin D. E., Strober W., Waldmann T. A. Ataxia-telangiectasia. Medicine (Baltimore) 1972 Jul;51(4):281–314. doi: 10.1097/00005792-197207000-00002. [DOI] [PubMed] [Google Scholar]
  22. Miller K. G., Liu L. F., Englund P. T. A homogeneous type II DNA topoisomerase from HeLa cell nuclei. J Biol Chem. 1981 Sep 10;256(17):9334–9339. [PubMed] [Google Scholar]
  23. Mohamed R., Pal Singh S., Kumar S., Lavin M. F. A defect in DNA topoisomerase II activity in ataxia-telangiectasia cells. Biochem Biophys Res Commun. 1987 Nov 30;149(1):233–238. doi: 10.1016/0006-291x(87)91629-9. [DOI] [PubMed] [Google Scholar]
  24. Morris C., Mohamed R., Lavin M. F. DNA replication and repair in ataxia telangiectasia cells exposed to bleomycin. Mutat Res. 1983 Apr;112(2):67–74. doi: 10.1016/0167-8817(83)90011-1. [DOI] [PubMed] [Google Scholar]
  25. North G. Eukaryotic topoisomerases come into the limelight. Nature. 1985 Aug 1;316(6027):394–395. doi: 10.1038/316394a0. [DOI] [PubMed] [Google Scholar]
  26. Oxford J. M., Harnden D. G., Parrington J. M., Delhanty J. D. Specific chromosome aberrations in ataxia telangiectasia. J Med Genet. 1975 Sep;12(3):251–262. doi: 10.1136/jmg.12.3.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. PETERSON R. D., KELLY W. D., GOOD R. A. ATAXIA-TELANGIECTASIA. ITS ASSOCIATION WITH A DEFECTIVE THYMUS, IMMUNOLOGICAL-DEFICIENCY DISEASE, AND MALIGNANCY. Lancet. 1964 May 30;1(7344):1189–1193. doi: 10.1016/s0140-6736(64)91209-7. [DOI] [PubMed] [Google Scholar]
  28. Painter R. B., Young B. R. Radiosensitivity in ataxia-telangiectasia: a new explanation. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7315–7317. doi: 10.1073/pnas.77.12.7315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Paterson M. C., Smith B. P., Lohman P. H., Anderson A. K., Fishman L. Defective excision repair of gamma-ray-damaged DNA in human (ataxia telangiectasia) fibroblasts. Nature. 1976 Apr 1;260(5550):444–447. doi: 10.1038/260444a0. [DOI] [PubMed] [Google Scholar]
  30. Shiloh Y., Tabor E., Becker Y. Repair of potentially lethal and sublethal damage induced by neocarzinostatin in normal and ataxia-telangiectasia skin fibroblasts. Biochem Biophys Res Commun. 1983 Jan 27;110(2):483–490. doi: 10.1016/0006-291x(83)91175-0. [DOI] [PubMed] [Google Scholar]
  31. Smith P. J., Anderson C. O., Watson J. V. Predominant role for DNA damage in etoposide-induced cytotoxicity and cell cycle perturbation in human SV40-transformed fibroblasts. Cancer Res. 1986 Nov;46(11):5641–5645. [PubMed] [Google Scholar]
  32. Taylor A. M., Harnden D. G., Arlett C. F., Harcourt S. A., Lehmann A. R., Stevens S., Bridges B. A. Ataxia telangiectasia: a human mutation with abnormal radiation sensitivity. Nature. 1975 Dec 4;258(5534):427–429. doi: 10.1038/258427a0. [DOI] [PubMed] [Google Scholar]
  33. Wang J. C. DNA topoisomerases. Annu Rev Biochem. 1985;54:665–697. doi: 10.1146/annurev.bi.54.070185.003313. [DOI] [PubMed] [Google Scholar]
  34. Yang L., Wold M. S., Li J. J., Kelly T. J., Liu L. F. Roles of DNA topoisomerases in simian virus 40 DNA replication in vitro. Proc Natl Acad Sci U S A. 1987 Feb;84(4):950–954. doi: 10.1073/pnas.84.4.950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. de Wit J., Jaspers N. G., Bootsma D. The rate of DNA synthesis in normal human and ataxia telangiectasia cells after exposure to X-irradiation. Mutat Res. 1981 Jan;80(1):221–226. doi: 10.1016/0027-5107(81)90190-1. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES