A DNA endonuclease isolated from yeast nuclear extract

Bryant, Douglas W.; Haynes, Robert H.

doi:10.1139/o78-031

Cited by 12 publications

(4 citation statements)

References 8 publications

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“…Yeast redoxyendonudease differs from the other previously described yeast UV endonucleases (Bryant & Haynes, 1978b;Bekker et al, 1980) and AP endonucleases (Pinon, 1970 and salt requirements, and other properties.…”

Section: Discussionmentioning

confidence: 70%

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Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III

Gossett

Lee²,

Cunningham³

et al. 1988

Biochemistry

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A DNA repair endonuclease (redoxyendonuclease) was isolated from bakers' yeast (Saccharomyces cerevisiae). The enzyme has been purified by a series of column chromatography steps and cleaves OsO4-damaged, double-stranded DNA at sites of thymine glycol and heavily UV-irradiated DNA at sites of cytosine, thymine, and guanine photoproducts. The base specificity and mechanism of phosphodiester bond cleavage for the yeast redoxyendonuclease appear to be identical with those of Escherichia coli endonuclease III when thymine glycol containing, end-labeled DNA fragments of defined sequence are employed as substrates. Yeast redoxyendonuclease has an apparent molecular size of 38,000-42,000 daltons and is active in the absence of divalent metal cations. The identification of such an enzyme in yeast may be of value in the elucidation of the biochemical basis for radiation sensitivity in certain yeast mutants.

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Section: Discussionmentioning

confidence: 70%

“…* State University of New York, Albany. Akhmedov et al, 1982), although there are reports identifying other enzymes including uracil-DNA glycosylase (Crosby et al, 1981) and UV-specific endonucleases (Bryant & Haynes, 1978b;Bekker et al, 1980).…”

mentioning

confidence: 99%

Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III

Gossett

Lee²,

Cunningham³

et al. 1988

Biochemistry

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“…A Mg2+-independent RNase was described by Ohtaka et al (1963) and Nakao et al (1968), and a Mg2+-dependent exonuclease degrading RNA from the 5' end was characterized by Stevens (1978Stevens ( , 1979Stevens ( , 1980. Nuclear enzymes also have been reported in yeasts, namely, two RNase H's (Wyers et al, 1976a,b) and a DNase (Bryant & Haynes, 1978). The DNase, however, was present at such low activity that it may have arisen from contaminating membrane material, including mitochondrial membranes.…”

mentioning

confidence: 99%

Purification and characteristics of a mitochondrial endonuclease from the yeast Saccharomyces cerevisiae

Tigerstrom¹

1982

Biochemistry

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Saccharomyces cerevisiae contains a membrane-bound mitochondrial nuclease. The enzyme was purified nearly 500-fold from sphaeroplasts of the organism by differential centrifugation, differential solubilization, heparin-agarose chromatography, and gel filtration. A final specific activity of 98 mumol min-1 (mg of protein)-1 was obtained. The enzyme required further purification to achieve homogeneity. Two peaks of activity were obtained after gel filtration with apparent molecular weights of 140000 and 57000. Otherwise, these two components have nearly identical characteristics. Without detergent the enzyme is insoluble and has very low activity. Zwittergent 3-14 or Triton X-100 in the presence of KCl could be used to solubilize and activate the enzyme. A number of other detergents were much less effective in solubilizing or activating the nuclease. The enzyme requires Mg2+ for activity, and this can be replaced to some degree by Mn2+ but not by Ca2+ or Zn2+. It is most active at pH 6.5-7.0 and degrades the substrate to small oligonucleotides with 5'-phosphate ends. The relative rates of hydrolysis were 100 for poly(A), 31 for ssDNA, 19 for RNA, 2.1 for dsDNA, and less than or equal to 0.2 for poly(C). Under the assay conditions used the enzyme appears to constitute about 90% of the total nuclease activity of the cell. The enzyme is unstable, especially at neutral and alkaline pH.

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“…8,10,11,26,28]. Their pH optima vary between pH 5.0 and 7.8 and some are unstable at 50°C [4]. Some have been reported to produce oligonucleotides with 5¢-phosphate termini [22,25,28].…”

Section: Discussionmentioning

confidence: 99%

Degradation of DNA during the autolysis of Saccharomyces cerevisiae

Zhao

Fleet

2003

J IND MICROBIOL BIOTECHNOL

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The autolysis of yeast cells has practical implications in the production of fermented foods and beverages and flavourants for food processing. Protein and RNA degradation during yeast autolysis are well described but the fate of DNA is unclear. Yeast cells ( Saccharomyces cerevisiae) were autolysed by incubating suspensions at 30-60 degrees C (pH 7.0), and at pH 4.0-7.0 (40 degrees C) for 10-14 days. Up to 55% of total DNA was degraded, with consequent leakage into the extracellular environment of mainly 3'- and 5'-deoxyribonucleotides, and lesser amounts of polynucleotides. The rate and extent of DNA degradation, composition of the DNA degradation products and DNase activity were affected by temperature and pH. The highest amount of DNA degradation occurred at 40 degrees C and pH 7.0, where the highest DNase activity was recorded. DNase activity was lowest at 60 degrees C and pH 4.0, where the proportion of polynucleotides in the degradation products was higher.

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A DNA endonuclease isolated from yeast nuclear extract

Cited by 12 publications

References 8 publications

Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III

Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III

Purification and characteristics of a mitochondrial endonuclease from the yeast Saccharomyces cerevisiae

Degradation of DNA during the autolysis of Saccharomyces cerevisiae

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