Advertisement

In Vitro Correction of Cells from Patients with Mutagen Hypersensitivity

  • N. G. J. Jaspers
  • L. Roza
  • W. Vermeulen
  • A. Eker
  • R. D. F. M. Taalman
  • J. H. J. Hoeijmakers
  • D. Bootsma
Chapter

Summary

An important strategy in the study of mutant cells, such as those from patients with an inherited hypersensitivity to UV or X-rays, is based on the principle of correction. In this contribution various approaches of in vitro correction are illustrated with complementation analysis of ataxia telangiectasia (AT) and with the introduction of wild-type protein extracts or yeast photoreactivating enzyme into xeroderma pigmentosum (XP) cells.

The combined results from genetic complementation studies in X-ray sensitive patients using the radioresistant DNA synthesis as a marker indicate the existence of six different complementation groups. Four of these (AB, C, D and E) are represented by AT patients and two (V1 and V2) harbour patients with the “Nijmegen Breakage syndrome”. A seventh separate category is formed by a number of AT patients with a normal inhibition of DNA synthesis by X-rays.

In XP cells a transient correction of the defect was accomplished by micro-injection of cell-free extracts from wild-type sources. Some properties of the factor correcting XP cells from complementation group A are described.

Micro-injection was also used to introduce the yeast photo-reactivating enzyme (PRE) into cells. After light-induced removal of pyrimidine dimers in injected cells the levels of UV-induced unscheduled DNA synthesis (UV-UDS) decreased only slowly. Kinetic studies suggest, that the average processing time of dimer excision is less than 1 hour in normal cells and more than 3 hours in some XP complementation groups.

Keywords

Ataxia Telangiectasia Average Processing Time Nijmegen Breakage Syndrome NIJMEGEN Breakage Syndrome Patient Mutagen Hypersensitivity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Tanaka K., M.Sekiguchi and Y.Okada (1975) Restoration of UV- induced unscheduled DNA synthesis in xeroderma pigmentosum cells by the concomitant treatment with bacteriophage T4 endonuclease and HVJ (Sendai virus). Proc.Natl.Ac.Sci. 72, 4071–4075CrossRefGoogle Scholar
  2. 2.
    De Jonge, A.J.R., W.Vermeulen, W.Keijzer, J.H.J.Hoeijmakers and D.Bootsma (1985) Microinjection of micrococcus Luteus W-endonuclease restores UV-induced unscheduled DNA synthesis in cells of nine xeroderma pigmentosum complementation groups. Mutation Res. 150, 99–105Google Scholar
  3. 3.
    Bridges, B.A. and D.G.Harnden, eds (1982) Ataxia telangiectasia: a molecular link between cancer, neuropathology and immune deficiency. Wiley and Sons Publ.Cy. ChichesterGoogle Scholar
  4. 4.
    Gatti, R.A. and M.Swift, eds (1985) Ataxia telangiectasia: genetics, immunology and neuropathology of a degenerative disease of childhood. Kroc Foundation Symp. Series, vol. 19, Alan Liss Publ. Cy., New York.Google Scholar
  5. 5.
    McKinnon, P.J. and L.A.Burgoyne (1987) Hum.Genet. 79, 197–208Google Scholar
  6. 6.
    Painter, R.B. (1981) Radioresistant DNA synthesis: an intrinsic feature of ataxia telangiectasia. Mutation Res. 84, 183–189PubMedCrossRefGoogle Scholar
  7. 7.
    Jaspers, N.G.J., J.De Wit, M.Regulski and D.Bootsma (1982) Increased lethality and abnormal regulation of DNA replication in ataxia telangiectasia cells treated with carcinogenic agents. Cancer Res. 42, 335–341Google Scholar
  8. 8.
    Fiorilli, M., A.Antonelli, G.Russo, M.Crescenzi, M.Carbonari and P.Petrinelli (1985) Variant of ataxia telangiectasia with low level radiosensitivity. Hum.Genet. 70, 274–277PubMedCrossRefGoogle Scholar
  9. 9.
    Jaspers, N.G.J. and D.Bootsma (1982) Genetic heterogeneity in ataxia telangiectasia studied by somatic cell fusion. Proc.Natl.Ac.Sci. 79, 2641–2644CrossRefGoogle Scholar
  10. 10.
    Murnane, J.P. and R.B.Painter (1982) Complementation of the defects in DNA synthesis in irradiated and unirradiated ataxia telangiectasia cells. Proc.Natl.Ac.Sci. 79, 1960–1963Google Scholar
  11. 11.
    Jaspers, N.G.J., R.B.Painter, M.C.Paterson, C.Kidson and T.Inoue (1985) Genetic complementation analysis of ataxia telangiectasia. in: ref. nr. 4, pp. 147–162Google Scholar
  12. 12.
    Paterson, M.C., B.P.Smith, P.A.Knight and A.K.Anderson (1977) Ataxia telangiectasia: an inherited human disease involving radiosensitivity, malignancy and defective DNA repair. In: Research in photobiology, 12.Paterson, M.C., B.P.Smith, P.A.Knight and A.K.Anderson, ed) pp 207–218 Plenum Press New YorkGoogle Scholar
  13. 13.
    Jaspers, N.G.J., R.D.F.M.Taalman, C.Baan (1988) Patients with an inherited syndrome characterized by immunodeficien cy, microcephaly and chromosomal instability: genetic rela tionship to ataxia telangiectasia. Am.Jrl.Hum.Genet. in pressGoogle Scholar
  14. 14.
    Weemaes, C.M.R., T.W.J.Hustinx, J.M.J.C.Scheres, P. J. J. van Munster, J.A.J.M.Bakkeren and R.D.F.M.Taalman (1981) New chromosome instability disorder: the nijmegen breakage syndrome. Acta Paed.Scand. 70, 557–562Google Scholar
  15. 15.
    Conley, M.E., M.B.Spinner, B.S.Emanuel, P.C.Nowell and W.W.Nichols (1985) A chromosome breakage syndrome with profound immunodeficiency. Blood 67, 1251–1256Google Scholar
  16. 16.
    Seemanova, E., E.Passarge, D.Beneskova, D.Houstek, P.Kasal and M.Sevcikova (1985) Familial microcephaly with normal intelligence, immunodeficiency and risk for lymphoreticular malignancy: A new autosomal recessive disorder. Am.Jrl.Med. Genet. 20, 639–648Google Scholar
  17. 17.
    Wegner, R.D., M.Metzger, F.Hanefeld, N.G.J.Jaspers, C.Baan, K.Magdorf and K.Sperling (1988) Clin.Genet., in pressGoogle Scholar
  18. 18.
    Zwetsloot, J.C.M., A.P.Barbeiro, W.Vermeulen, H.M.Arthur, J.H.J.Hoeijmakers and C.Backendorf (1986) Microinjection of Escherichia coli uvrA,B,C and D proteins into fibroblasts of xeroderma pigmentosum groups A and C does not result in restoration of UV-induced unscheduled DNA synthesis. Muta tion Res. 166, 89–98CrossRefGoogle Scholar
  19. 19.
    Keyzer, W., A.Verkerk and D.Bootsma (1982) Phenotypic correction of the defect in xeroderma pigmentosum after fusion with isolated cytoplasts. Exp.Cell Res. 140, 119–125CrossRefGoogle Scholar
  20. 20.
    Vermeulen,W., P.Osseweijer, A. J. R. de Jonge and J.H.J.Hoeijmakers (1986) Transient correction of excision repair defects in fibroblasts of nine xeroderma pigmentosum compie mentation groups by microinjection of crude human cell extracts. Mutat. Res. 165, 199–206Google Scholar
  21. 21.
    J.H.J.Hoeijmakers (1987) Characterization of genes and proteins involved in excision repair of human cells. Jrl.Ceil Sci. suppl. 6, 111–125Google Scholar
  22. 22.
    Legerski, R.J., D.B.Brown, C.A.Peterson and D.L.Robberson (1984) Transient complementation of xeroderma pigmentosum cells by microinjection with poly(A)+RNA. Proc.Natl.Ac.Sci. 81, 5676–5679CrossRefGoogle Scholar
  23. 23.
    Yamaizumi, M., T.Sugano, H.Asahina, Y.Okada and T.Uchida (1986) Microinjection of partially purified protein factor restores DNA damage specifically in group A of xeroderma pigmentosum cells. Proc.Natl.Ac.Sci. 83, 1476–1479CrossRefGoogle Scholar
  24. 24.
    Zwetsloot, J.C.M., J.H.J.Hoeijmakers, W.Vermeulen, A.P.M. Eker and D.Bootsma (1986) Unscheduled DNA synthesis in xeroderma pigmentosum cells after microinjection of yeast photoreactivating enzyme. Mutation Res, 165, 109–115Google Scholar
  25. 25.
    Zwetsloot, J.C.M., W.Vermeulen, J.H.J.Hoeijmakers, A.Yasui, A.P.M.Eker and D.Bootsma (1985) Microinjected photoreactivating enzymes from Anacystis and Saccharomyces monomerize dimers in chromatin of human cells. Mutat.Res. l46, 71–77Google Scholar
  26. Paterson, M.C., M.V.Middlestadt, S.J.McFarlane, N.E.Gentner, M.Weinfeld and A.P.M.Eker (1987) Molecular evidence for cleavage of intradimer phosphodiester linkage as a novel step in excision repair of cyclobutyl pyrimidine photodimers in cultured human cells. Jrl. Cell Sci.suppl.6, 161–176Google Scholar

Copyright information

© Springer Science+Business Media New York 1989

Authors and Affiliations

  • N. G. J. Jaspers
    • 1
  • L. Roza
    • 2
  • W. Vermeulen
    • 1
  • A. Eker
    • 1
  • R. D. F. M. Taalman
    • 3
  • J. H. J. Hoeijmakers
    • 1
  • D. Bootsma
    • 1
  1. 1.Laboratory of Cell Biology and GeneticsErasmus UniversityRotterdamThe Netherlands
  2. 2.Medical Biological LaboratoryRijswijkThe Netherlands
  3. 3.Department of Human GeneticsCatholic Univ.NijmegenThe Netherlands

Personalised recommendations