Purification and characterization of a uracil-DNA glycosylase from the yeast, saccharomyces cerevisiae

Bill Crosby, Louise Prakash, Howard Davis, David C. Hinkle

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

An activity which releases free uracil from bacteriophage PBS1 DNA has been purified over 10,000 fold from extracts of Saccharornyces cerevisiae. The enzyme is active on both native and denatured PBSl UNA and is active in the absence of divalent cation, and in the presence of 1 mM EDTA. The enzyme has a native molecular weight of 27,800 as estimated by glycerol gradient centrifugation and gelfiltration. Enzyme activity has been recovered after denaturation in SDS and electrophoresis in an SDS polyacrylamide gel. This analysis suggests that the enzyme consists of a single polypeptide chain of about 27,000 daltons. Normal levels of uracil-DNA glycosylase activity were found in partially purified extracts of the nitrous-acid sensitive rad18-2 mutant of yeast.

Original languageEnglish (US)
Pages (from-to)5797-5810
Number of pages14
JournalNucleic Acids Research
Volume9
Issue number21
DOIs
StatePublished - Nov 11 1981
Externally publishedYes

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Uracil-DNA Glycosidase
Saccharomyces Cerevisiae
Purification
Yeast
Saccharomyces cerevisiae
Enzymes
DNA
Yeasts
Nitrous Acid
Denaturation
Bacteriophages
Uracil
Centrifugation
Polypeptides
Ethylenediaminetetraacetic acid
Divalent Cations
Enzyme activity
Polyacrylates
Electrophoresis
Glycerol

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Purification and characterization of a uracil-DNA glycosylase from the yeast, saccharomyces cerevisiae. / Crosby, Bill; Prakash, Louise; Davis, Howard; Hinkle, David C.

In: Nucleic Acids Research, Vol. 9, No. 21, 11.11.1981, p. 5797-5810.

Research output: Contribution to journalArticle

Crosby, Bill ; Prakash, Louise ; Davis, Howard ; Hinkle, David C. / Purification and characterization of a uracil-DNA glycosylase from the yeast, saccharomyces cerevisiae. In: Nucleic Acids Research. 1981 ; Vol. 9, No. 21. pp. 5797-5810.
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