Purification and characterization of a uracil-DNA glycosylase from the yeast, Saccharomyces cerevisiae
- 1 January 1981
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 9 (21), 5797-5810
- https://doi.org/10.1093/nar/9.21.5797
Abstract
An activity which releases free uracil from bacteriophage PBS1 DNA has been purified over 10,000 fold from extracts of Saccharomyces cerevisiae. The enzyme is active on both native and denatured PBS1 DNA and is active in the absence of divalent cation, and in the presence of 1 mM EDTA. The enzyme has a negative molecular weight of 27,800 as estimated by glycerol gradient centrifugation and gel filtration. 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.Keywords
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