Identification of functional domains within the RAD1·RAD10 repair and recombination endonuclease of Saccharomyces cerevisiae

Karl Rodriguez, Zhigang Wang, Errol C. Friedberg, Alan E. Tomkinson

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Saccharomyces cerevisiae rad1 and rad10 mutants are unable to carry out nucleotide excision repair and are also defective in a mitotic intrachromosomal recombination pathway. The products of these genes are subunits of an endonuclease which recognizes DNA duplex/single-strand junctions and specifically cleaves the 3' single-strand extension at or near the junction. It has been suggested that such junctions arise as a consequence of DNA lesion processing during nucleotide excision repair and the processing of double-strand breaks during intrachromosomal recombination. In this study we show that the RAD1·RAD10 complex also cleaves a more complex junction structure consisting of a duplex with a protruding 3' single-strand branch that resembles putative recombination intermediates in the RAD1·RAD10-mediated single-strand annealing pathway of mitotic recombination. Using monoclonal antibodies, we have identified two regions of RAD1 that are required for the cleavage of duplex/single-strand junctions. These reagents also inhibit nucleotide excision repair in vitro, confirming the essential role of the RAD1·RAD10 endonuclease in this pathway.

Original languageEnglish (US)
Pages (from-to)20551-20558
Number of pages8
JournalJournal of Biological Chemistry
Issue number34
StatePublished - Sep 5 1996


ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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