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

Yvette Habraken, Patrick Sung, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

110 Citations (Scopus)

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 pigmentosum (XP) in humans1,2. The RAD2 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 double-stranded 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.

Original languageEnglish (US)
Pages (from-to)365-368
Number of pages4
JournalNature
Volume366
Issue number6453
StatePublished - Nov 25 1993

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Single-Stranded DNA
Deoxyribonuclease I
DNA Repair
Yeasts
Xeroderma Pigmentosum
Endonucleases
Eukaryota
Genes
DNA
Saccharomyces cerevisiae Proteins
Circular DNA
Saccharomyces
Proteins
Magnesium
Radiation
Neoplasms

ASJC Scopus subject areas

  • General

Cite this

Yeast excision repair gene RAD2 encodes a single-stranded DNA endonuclease. / Habraken, Yvette; Sung, Patrick; Prakash, Louise; Prakash, Satya.

In: Nature, Vol. 366, No. 6453, 25.11.1993, p. 365-368.

Research output: Contribution to journalArticle

Habraken, Yvette ; Sung, Patrick ; Prakash, Louise ; Prakash, Satya. / Yeast excision repair gene RAD2 encodes a single-stranded DNA endonuclease. In: Nature. 1993 ; Vol. 366, No. 6453. pp. 365-368.
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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 pigmentosum (XP) in humans1,2. The RAD2 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 double-stranded 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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N2 - 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 pigmentosum (XP) in humans1,2. The RAD2 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 double-stranded 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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