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Repair of 8-methoxypsoralen photoinduced cross-links in yeast

Analysis by alkaline step-elution and electron microscopy

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Summary

The repair of interstrand cross-links induced by 8-methoxypsoralen plus UVA (365 nm) radiation DNA was analyzed in diploid strains of the yeast Saccharomyces cerevisiae. The strains employed were the wild-type D7 and derivatives homozygous for the rad18-1 or the rad3-12 mutation. Alkaline step-elution and electron microscopy were performed to follow the process of induction and removal of photoinduced crosslinks. In accordance with previous reports, the D7 rad3-12 strain failed to remove the induced lesions and could not incise cross-links. The strain D7 rad18-1 was nearly as efficient in the removal of 8-MOP photoadducts after 2 h of post-treatment incubation as the D7 RAD+ wild-type strain. However, as demonstrated by alkaline step-elution and electron microscopic analysis, the first incision step at DNA cross-links was three times more effective in D7 rad18-1 than in D7 RAD+. This is consistent with the hypothesis that the RAD18 gene product is involved in the filling of gaps resulting from persistent non-informational DNA lesions generated by the endonucleolytic processing of DNA cross-links. Absence of this gene product may lead to extensive strand breakage and decreased recognition of such lesions by structural repair systems.

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Authors and Affiliations

  1. Istituto di Mutagenesi e Differenziamento C.N.R., I-56100, Pisa, Italy

    E. Cundari

  2. Institut Curie - Biologic, CNRS URA 1292, 26 rue d'Ulm, F-75231, Paris Cedex 05, France

    M. Dardalhon, S. Rousset & D. Averbeck

Authors
  1. E. Cundari
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  2. M. Dardalhon
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  3. S. Rousset
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  4. D. Averbeck
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Communicated by W. Gajewski

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Cundari, E., Dardalhon, M., Rousset, S. et al. Repair of 8-methoxypsoralen photoinduced cross-links in yeast. Molec. Gen. Genet. 228, 335–344 (1991). https://doi.org/10.1007/BF00260625

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  • DOI: https://doi.org/10.1007/BF00260625

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