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Yeast excision repair gene RAD2 encodes a single-stranded DNA endonuclease

Nature volume 366, pages 365–368 (1993)Cite this article

Abstract

IN eukaryotes nucleotide excision repair of DNA damaged by ultraviolet radiation requires several gene products; defects in this process result in the cancer-prone syndrome xeroderma pigmento-sum (XP) in humans1,2. TheRAD2 gene is one of at least seven genes indispensable for excision repair in the yeast Saccharomyces cerevisiae2, and its encoded protein shares remarkable homology with the XP group-G gene product3. Here we overproduce the RAD2-encoded protein in S. cerevisiae, purify it to near homogeneity, and show that RAD2 protein in the presence of magnesium degrades circular single-stranded DNA. The RAD2 endonuclease is specific for single-stranded DNA as it does not act on doublestranded DNA. Given the absolute requirement for RAD2 in the incision step of excision repair, our findings directly implicate RAD2 protein and its human homologue XPG protein as a catalytic component that incises the damaged DNA strand during excision repair. Furthermore, our results indicate that eukaryotes probably employ two distinct endonuclease activities to mediate the dual incision at the damage site.

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Author notes

  1. Yvette Habraken, Patrick Sung, Louise Prakash & Satya Prakash

    Present address: UTMB, Sealy Center for Molecular Science, Medical Research Building, Route J61, 11th Street and Mechanic, Galveston, Texas, 77555-1061, USA

Authors and Affiliations

  1. Department of Biophysics, University of Rochester, Rochester, New York, 14642, USA

    Yvette Habraken & Louise Prakash

  2. Department of Biology, University of Rochester, Rochester, New York, 14642, USA

    Patrick Sung & Satya Prakash

Authors
  1. Yvette Habraken
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  2. Patrick Sung
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  3. Louise Prakash
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  4. Satya Prakash
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Habraken, Y., Sung, P., Prakash, L. et al. Yeast excision repair gene RAD2 encodes a single-stranded DNA endonuclease. Nature 366, 365–368 (1993). https://doi.org/10.1038/366365a0

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