Yeast DNA-repair gene RAD14 encodes a zinc metalloprotein with affinity for ultraviolet-damaged DNA

Sami N. Guzder, Patrick Sung, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

86 Scopus citations

Abstract

Xeroderma pigmentosum (XP) patients suffer from a high incidence of skin cancers due to a defect in excision repair of UV light-damaged DNA. Of the seven XP complementation groups, A-G, group A represents a severe and frequent form of the disease. The Saccharomyces cerevisiae RAD14 gene is a homolog of the XP-A correcting (XPAC) gene. Like XP-A cells, rad14-null mutants are defective in the incision step of excision repair of UV-damaged DNA. We have purified RAD14 protein to homogeneity from extract of a yeast strain genetically tailored to overexpress RAD14. As determined by atomic emission spectroscopy, RAD14 contains one zinc atom. We also show in vitro that RAD14 binds zinc but does not bind other divalent metal ions. In DNA mobility-shift assays, RAD14 binds specifically to UV-damaged DNA. Removal of cyclobutane pyrimidine dimers from damaged DNA by enzymatic photoreactivation has no effect on binding, strongly suggesting that RAD14 recognizes pyrimidine(6-4)pyrimidone photoproduct sites. These findings indicate that RAD14 functions in damage recognition during excision repair.

Original languageEnglish (US)
Pages (from-to)5433-5437
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number12
DOIs
StatePublished - Jun 15 1993
Externally publishedYes

Keywords

  • DNA damage-binding protein
  • Saccharomyces cerevisiae
  • Xeroderma pigmentosum

ASJC Scopus subject areas

  • General

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