Defective DNA cross-link removal in Chinese hamster cell mutants hypersentitive to bifunctional alkylating agents

C. A. Hoy, L. H. Thompson, C. L. Mooney, E. P. Salazar

Research output: Contribution to journalArticle

222 Scopus citations

Abstract

DNA repair-deficient mutants from five genetic complementation groups isolated previously from Chinese hamster cells were assayed for survival after exposure to the bifunctional alkylating agents mitocmycin C or diepoxybutane. Groups 1, 3, and 5 exhibited 1.6- to 3-fold hypersensitivity compared to the wild-type cells, whereas Groups 2 and 4 exhibited extraordinary hypersensitivity (30- to 90-fold). Mutants from Groups 1 and 2 were exposed to 22 other bifunctional alkylating agents in a rapid assay that compared cytotoxicity of the mutants to the wild-type parental strain, AA8. With all but two of the compounds, the Group 2 mutant (UV4) was 15- to 60-fold more sensitive than AA8 or the Group 1 mutant (UV5). UV4 showed only 6-fold hypersensitivity to quinacrine mustard. Alkaline elution measurements showed that this compound produced few DNA interstrand cross-links but numerous strand breaks that were revealed by proteinase treatment. Therefore, the extreme hypersensitivity of mutants from Groups 2 and 4 appeared specific for compounds the main cytotoxic lesions of which were DNA cross-links. Mutant UV5 was only 1- to 4-fold hypersensitive to all the compounds. Repair kinetics of DNA interstrand cross-link production and removal was measured by alkaline elution for AA8 and mutants UV4 and UV5 after exposure to diepoxybutane. Although the initial number of cross-links was similar for the three cell lines, during 24-h incubation, the efficiency of removal of cross-links was lowest in UV4 and intermediate in UV5. These results suggest that the different levels of sensitivity of the five complementation groups to bifunctional alkylation damage are specifically related to different efficiencies of DNA cross-link removal. The phenotype of hypersensitivity of both UV radiation and cross-link damage exhibited by the mutants in Groups 2 and 4 appears to differ from those of the known human DNA repair syndromes.

Original languageEnglish (US)
Pages (from-to)1737-1743
Number of pages7
JournalCancer Research
Volume45
Issue number4
StatePublished - 1985
Externally publishedYes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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    Hoy, C. A., Thompson, L. H., Mooney, C. L., & Salazar, E. P. (1985). Defective DNA cross-link removal in Chinese hamster cell mutants hypersentitive to bifunctional alkylating agents. Cancer Research, 45(4), 1737-1743.