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Formation and fate of cross-links induced by polyfunctional anticancer drugs in yeast

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Summary

A method to detect low levels of interstrand cross-links in DNA of Saccharomyces cerevisiae is described. Isopycnic ultracentrifugation of alkali-treated, unpurified Eaton press homogenates allows the detection of less than one cross-link per yeast chromosome. Efficient separation of single-and double-stranded DNA requires low cell density and addition of glycerol during homogenization. Using a yeast strain defective in excision repair, a dose dependent formation of interstrand cross-links after treatment of cells with biological doses of nitrogen mustard. Triaziquone and Chloramubil could be demonstrated. The most powerful of these alkylating agents is Triaziquone: half of the DNA molecules are shown to be cross-linked after a 12 min exposure to 9×10-9 g/ml of the drug. The cross-linking reaction continues after excessive alkylating agent is removed. After having reached a maximum the fraction continues after excessive alkylating agent is removed. After having reached a maximum the fraction of renaturable DNA decreases upon further incubation. The speed of this “after-reaction” depends on temperature: 48 h after the end of treatment renaturability of DNA has almost completely disappeared when cells are kept at 36° C.

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  1. Fachbereiche Biochemie/Pharmazie and Biologie, J.W. Goethe-Universität, D-6000, Frankfurt/Main, Federal Republic of Germany

    Reinhard Fleer & Martin Brendel

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  1. Reinhard Fleer
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  2. Martin Brendel
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Communicated by F. Kaudewitz

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Fleer, R., Brendel, M. Formation and fate of cross-links induced by polyfunctional anticancer drugs in yeast. Molec. Gen. Genet. 176, 41–52 (1979). https://doi.org/10.1007/BF00334294

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  • DOI: https://doi.org/10.1007/BF00334294

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