Skip to main content
Log in

Analysis of bromodeoxyuridine-induced single and twin sister chromatid exchanges in tetraploid Chinese hamster ovary cells

  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

Culture of cells in high exogenous levels (>10−4 M) of bromodeoxyuridine (BrdUrd) or thymidine will increase the baseline sister chromatid exchange (SCE) frequency. The effect is thought to be related to the balance of the DNA precursors thymidine and deoxycytidine. Exogenous addition of deoxycytidine will reverse this effect. Single and twin SCEs were analysed in Colcemid-induced tetraploid Chinese hamster ovary cells exposed to different concentrations of BrdUrd to determine at what stage SCEs are induced by high levels of BrdUrd. In cells exposed to low concentrations of BrdUrd (10−5 M), equal numbers of SCEs were induced in each of the two cell cycles. With increasing concentrations of BrdUrd (10−4 to 2×10−4 M), SCE frequency increased in both cell cycles, but far more SCEs were induced in the second cell cycle. Deoxycytidine (2×10−4 M) reduced the frequency of SCEs primarily by reducing the frequency of SCEs induced in the second cell cycle. Treatment with 3-aminobenzamide (3AB), a potent inhibitor of poly(ADP-ribose) polymerase, produced effects similar to exposure to high levels of BrdUrd including inducing SCEs in the second replication cycle. This suggests a similar mechanism of action. Deoxycytidine had no effect on 3AB-induced SCEs, however, and there was no interaction between 3AB and high exogenous levels of BrdUrd in SCE induction. Thus these two agents probably act through different mechanisms.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Cleaver JE (1984) Differential toxicity of 3-aminobenzamide to wild-type and 6-thioguanine-resistant Chinese hamster cells by interference with pathways of purine biosynthesis. Mutat Res 131:123–127

    Google Scholar 

  • Davidson, RL, Kaufmann ER, Dougherty CP, Ouellette AM, Di-Folco CM, Latt SA (1980) Induction of sister chromatid exchanges by BUdR is largely independent of the BUdR content of DNA. Nature 284:74–76

    Google Scholar 

  • Durkacz BW, Omidiji O, Gray DA, Shall S (1980) (ADP-ribose)n participates in DNA excision repair. Nature 283:593–596

    Google Scholar 

  • Durrant LG, Boyle JM (1982) Potentiation of cell killing by inhibitors of poly(ADP-ribose) polymerase in four rodent cell lines exposed to N-methyl-N-nitrosourea or UV light. Chem Biol Interact 38:325–338

    Google Scholar 

  • Goto K, Akematsu T, Shimazu H, Sugiyama T (1975) Simple differential Giemsa staining of sister chromatids after treatment with photosensitive dyes and exposure to light and the mechanism of staining. Chromosoma 53:223–230

    Google Scholar 

  • Heartline MW, O'Neill JP, Preston RJ (1983) SCE induction is proportional to substitution in DNA for thymidine by CldU and BrdU. Mutat Res 107:103–104

    Google Scholar 

  • Heddle JA (1969) Influence of false twins on the ratios of twin and single sister chromatid exchanges. J Theor Biol 22:151–162

    Google Scholar 

  • Krepinsky AB, Heddle JA, German J (1979) Sensitivity of Bloom's Syndrome lymphocytes to ethyl methanesulfonate. Hum Genet 50:151–156

    Google Scholar 

  • Latt SA (1973) Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc Natl Acad Sci USA 70:3395–3399

    Google Scholar 

  • Linnainmaa K, Wolff S (1982) Sister chromatid exchanges induced by short-lived monoadducts produced by the bifunctional agents mitomycin C and 8-methoxypsoralen. Environ Mutagen 4:239–247

    Google Scholar 

  • Mazrimas JA, Stetka DG (1978) Direct evidence for the role of incorporated BUdR in induction of sister chromatid exchanges. Exp Cell Res 117:23–30

    Google Scholar 

  • Meuth M (1981a) The role of deoxynucleoside triphosphate pools in the cytotoxic and mutagenic effects of DNA alkylating agents. Somatic Cell Genet 7:89–102

    Google Scholar 

  • Meuth M (1981b) Sensitivity of a mutator gene in Chinese hamster ovary cells to deoxynucleoside triphosphate pool alterations. Mol Cell Biol 1:652–660

    Google Scholar 

  • Milam K, Cleaver JE (1984) Inhibitors of poly(ADP-ribose) synthesis also affect other metabolic processes. Science 223:589–591

    Google Scholar 

  • Morgan WF, Wolff S (1984) Induction of sister-chromatid exchange by 3-aminobenzamide is independent of bromodeoxyuridine. Cytogenet Cell Genet 38:34–38

    Google Scholar 

  • Natarajan AT, Csukas I, van Zeeland AA (1981) Contribution of incorporated 5-bromodeoxyuridine in DNA to the frequencies of sister chromatid exchanges by inhibitors of poly(ADPribose)-polymerase. Mutat Res 84:125–132

    Google Scholar 

  • Oikawa A, Tohda H, Kanai M, Miwa M, Sugimura T (1980) Inhibitors of poly(adenosine diphosphate ribose) polymerase induce sister chromatid exchanges. Biochem Biophys Res Commun 97:1311–1316

    Google Scholar 

  • Perry P, Wolff S (1974) New Giemsa method for the differential staining of sister chromatids. Nature 251:156–158

    Google Scholar 

  • Perry PE (1983) Induction of sister-chromatid exchanges (SCEs) by thymidine and the potentiation of mutagen-induced SCEs in Chinese hamster ovary cells. Mutat Res 109:219–229

    Google Scholar 

  • Peterson AR, Landolph Jr, Peterson H, Heidelberger C (1978) Mutagenesis of Chinese hamster cells is facilitated by thymidine and deoxycytidine. Nature 276:508–510

    Google Scholar 

  • Schwartz JL, Banda MJ, Wolff S (1982) 12-O-tetradecanoylphorbol-13-acetate (TPA) induces sister chromatid exchanges and delays in cell progression in Chinese hamster ovary and human cell lines. Mutat Res 92:393–409

    Google Scholar 

  • Schwartz JL, Weichselbaum RR (1985) Antipain mediated suppression of sister chromatid exchanges induced by an inhibitor of poly(ADP-ribose) polymerase. Environ Mutagen 7:703–709

    Google Scholar 

  • Schwartz JL, Morgan WF, Brown-Lindquist P, Afzal V, Weichselbaum RR, Wolff S (1985a) Comutagenic effects of 3-aminobenzamide in Chinese hamster ovary cells. Cancer Res 45:1556–1559

    Google Scholar 

  • Schwartz JL, Morgan WF, Weichselbaum RR (1985b) Different efficiencies of interaction between 3-aminobenzamide and various monofunctional alkylating agents in the induction of sister chromatid exchanges. Carcinogenesis 6:699–704

    Google Scholar 

  • Shall S (1984) ADP-ribose in DNA repair. A new component of DNA excision repair. Adv Radiat Biol 11:2–64

    Google Scholar 

  • Shiraishi Y, Freeman AI, Sandberg AA (1976) Increased sister chromatid exchange in bone marrow and blood cells from Bloom's syndrome. Cytogenet Cell Genet 17:162–173

    Google Scholar 

  • Shiraishi Y, Yosida TH, Sandberg AA (1982) Analysis of single and twin sister chromatid exchanges in endoreduplicated normal and Bloom's syndrome B-lymphoid cells. Chromosoma 87:1–8

    Google Scholar 

  • Shiraishi Y, Tanaka Y, Kato M, Miwa M, Sugimura T (1983) Effect of poly (ADP-ribose) polymerase inhibitors on the frequency of sister-chromatid exchanges in Bloom syndrome cells. Mutat Res 122:223–228

    Google Scholar 

  • Shiraishi Y, Yosida TH, Sandberg AA (1985) Inhibition of bromodeoxyuridine associated sister chromatid exchanges in Bloom's syndrome cells with cycloheximide. Cancer Genet Cytogent 17:43–54

    Google Scholar 

  • Stetka DG, Carrano AV (1977) The interaction of Hoechst 33258 and BrdU substituted DNA in the formation of sister chromatid exchanges. Chromosoma 63:21–31

    Google Scholar 

  • Taylor JH (1958) Sister chromatid exchanges in tritium-labeled chromosomes. Genetics 43:515–529

    Google Scholar 

  • Terasima T, Tolmach LJ (1961) Changes in x-ray sensitivity of HeLa cells during the division cycle. Nature 190:1210–1211

    Google Scholar 

  • Wolff S, Perry P (1975) Insights on chromosome structure from sister chromatid exchange ratios and the lack of both isolabelling and heterolabelling as determined by the FPG technique. Exp Cell Res 93:23–30

    Google Scholar 

  • Wolff S (1978) Chromosomal effects of mutagenic carcinogens and the nature of the lesions leading to sister chromatid exchange. In: Evans HJ, Lloyd DC (eds) Mutagen-induced chromosome damage in man. Edinburgh University Press, pp 208–215

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schwartz, J.L. Analysis of bromodeoxyuridine-induced single and twin sister chromatid exchanges in tetraploid Chinese hamster ovary cells. Chromosoma 93, 409–412 (1986). https://doi.org/10.1007/BF00285822

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00285822

Keywords

Navigation