A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae

Richard Kostriken, Jeffrey N. Strathern, Amar J S Klar, James B. Hicks, Fred Heffron

Research output: Contribution to journalArticle

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Abstract

We have detected two site-specific endonucleases in strains of Saccharomyces cerevisiae. One endonuclease, which we call YZ endo, is present only in yeast strains that are undergoing mating-type interconversion. The site at which YZ endo cleaves corresponds to the in vivo double-strand break occurring at the mating-type locus in,yeast undergoing mating-type interconversion. YZ endo generates a site-specific double-strand break having 4-base 3′ extensions terminating in 3′ hydroxyl groups. The site of cleavage occurs in the Z1 region near the YZ junction of the mating-type locus. Mutant mating-type loci known to decrease the frequency of mating-type interconversion are correspondingly poor substrates for YZ endo in vitro. In vitro analysis of a number of such altered recognition sites has delimited the sequences required for cleavage. The molecular genetics of mating-type interconversion is discussed in the context of this endonucleolytic activity. The second endonuclease, which we refer to as Sce II, is present in all strains of S. cerevisiae we have examined. The cleavage site of Sce II has been determined and proves to be unrelated to the cleavage site of YZ endo.

Original languageEnglish (US)
Pages (from-to)167-174
Number of pages8
JournalCell
Volume35
Issue number1
DOIs
StatePublished - 1983
Externally publishedYes

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Endonucleases
Yeast
Saccharomyces cerevisiae
Yeasts
Hydroxyl Radical
Molecular Biology
Substrates
In Vitro Techniques

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Kostriken, R., Strathern, J. N., Klar, A. J. S., Hicks, J. B., & Heffron, F. (1983). A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae. Cell, 35(1), 167-174. https://doi.org/10.1016/0092-8674(83)90219-2

A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae. / Kostriken, Richard; Strathern, Jeffrey N.; Klar, Amar J S; Hicks, James B.; Heffron, Fred.

In: Cell, Vol. 35, No. 1, 1983, p. 167-174.

Research output: Contribution to journalArticle

Kostriken, R, Strathern, JN, Klar, AJS, Hicks, JB & Heffron, F 1983, 'A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae', Cell, vol. 35, no. 1, pp. 167-174. https://doi.org/10.1016/0092-8674(83)90219-2
Kostriken, Richard ; Strathern, Jeffrey N. ; Klar, Amar J S ; Hicks, James B. ; Heffron, Fred. / A site-specific endonuclease essential for mating-type switching in Saccharomyces cerevisiae. In: Cell. 1983 ; Vol. 35, No. 1. pp. 167-174.
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AB - We have detected two site-specific endonucleases in strains of Saccharomyces cerevisiae. One endonuclease, which we call YZ endo, is present only in yeast strains that are undergoing mating-type interconversion. The site at which YZ endo cleaves corresponds to the in vivo double-strand break occurring at the mating-type locus in,yeast undergoing mating-type interconversion. YZ endo generates a site-specific double-strand break having 4-base 3′ extensions terminating in 3′ hydroxyl groups. The site of cleavage occurs in the Z1 region near the YZ junction of the mating-type locus. Mutant mating-type loci known to decrease the frequency of mating-type interconversion are correspondingly poor substrates for YZ endo in vitro. In vitro analysis of a number of such altered recognition sites has delimited the sequences required for cleavage. The molecular genetics of mating-type interconversion is discussed in the context of this endonucleolytic activity. The second endonuclease, which we refer to as Sce II, is present in all strains of S. cerevisiae we have examined. The cleavage site of Sce II has been determined and proves to be unrelated to the cleavage site of YZ endo.

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