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UV-induced endonuclease III-sensitive sites at the mating type loci inSaccharomyces cerevisiae are repaired by nucleotide excision repair: RAD7 and RAD16 are not required for their removal fromHMLα

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Abstract

Ultraviolet irradiation of DNA induces cyclobutane pyrimidine dimers (CPDs), 6-4′-(pyrimidine 2′-one) pyrimidines and pyrimidine hydrates. The dimer is the major photoproduct, and is specifically recognized by endonuclease V of phage T4. Pyrimidine hydrates represent a small fraction of the total photoproducts, and are substrates for endonuclease III ofEscherichia coli. We used these enzymes to follow the fate of their substrates in the mating type loci ofSaccharomyces cerevisiae. In a RAD strain, CPDs in the transcriptionally activeMATα locus are preferentially repaired relative to the inactiveHMLα locus, whilst repair of endonuclease III-sensitive sites is not preferential. Therad1, 2, 3 and4 mutants, which lack factors that are essential for the incision step of nucleotide excision repair (NER), repair neither CPDs nor endonuclease III-sensitive sites, clearly showing that these lesions are repaired by the NER pathway. Previously it had been shown that the products of theRAD7 andRAD16 genes are required for the NER of CPDs from theHMLα locus. We show that, in the same locus, these gene products are not needed for removal of endonuclease III-sensitive sites by the same mechanism. This indicates that the components required for NER differ depending on either the type of lesion encountered or on the specific location of the lesion within the genome.

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Authors and Affiliations

  1. School of Biological Sciences, University College Swansea, Singleton Park, SA2 8PP, Swansea, UK

    S. H. Reed & R. Waters

  2. Centre National de la Recherche Scientifique, URA 147, Institut Gustave-Roussy, 94800, Villejuif, France

    S. Boiteux

Authors
  1. S. H. Reed
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  2. R. Waters
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  3. S. Boiteux
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Communicated by B. J. Kilbey

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Reed, S.H., Waters, R. & Boiteux, S. UV-induced endonuclease III-sensitive sites at the mating type loci inSaccharomyces cerevisiae are repaired by nucleotide excision repair: RAD7 and RAD16 are not required for their removal fromHMLα . Molec. Gen. Genet. 250, 505–514 (1996). https://doi.org/10.1007/BF02174039

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