Renaturation of DNA by a Saccharomyces cerevisiae protein that catalyzes homologous pairing and strand exchange

W. D. Heyer, D. H. Evans, R. D. Kolodner

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

A protein from mitotic Saccharomyces cerevisiae cells that catalyzes homologous pairing and strand exchange was analyzed for the ability to catalyze other related reactions. The protein was capable of renaturing complementary single-stranded DNA as evidenced by S1 nuclease assays and analysis of the reaction products by agarose gel electrophoresis and electron microscopy. Incubation of the yeast protein with complementary single-stranded DNA resulted in the rapid formation of large aggregates which did not enter agarose gels. These aggregates contained many branched structures consisting of both single-stranded and double-stranded DNA. These reactions required stoichiometric amounts of protein but showed no ATP requirement. The protein formed stable complexes with both single-stranded and double-stranded DNA, showing a higher affinity for single-stranded DNA. The binding to single-stranded DNA resulted in the formation of large protein:DNA aggregates. These aggregates were also formed in strand-exchange reactions and contained both substrate and product DNAs. These results demonstrate that the S. cerevisiae strand-exchange protein shares additional properties with the Escherichia coli recA protein which, by analogy, gives further indication that it might be implicated in homologous recombination.

Original languageEnglish (US)
Pages (from-to)15189-15195
Number of pages7
JournalJournal of Biological Chemistry
Volume263
Issue number29
StatePublished - 1988
Externally publishedYes

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Saccharomyces cerevisiae Proteins
Single-Stranded DNA
DNA
Proteins
Complementary DNA
Rec A Recombinases
Yeast
Sepharose
Fungal Proteins
Agar Gel Electrophoresis
Escherichia coli Proteins
Homologous Recombination
Gels
Saccharomyces cerevisiae
Electron Microscopy
Adenosine Triphosphate
Electrophoresis
Reaction products
Escherichia coli
Electron microscopy

ASJC Scopus subject areas

  • Biochemistry

Cite this

Renaturation of DNA by a Saccharomyces cerevisiae protein that catalyzes homologous pairing and strand exchange. / Heyer, W. D.; Evans, D. H.; Kolodner, R. D.

In: Journal of Biological Chemistry, Vol. 263, No. 29, 1988, p. 15189-15195.

Research output: Contribution to journalArticle

Heyer, W. D. ; Evans, D. H. ; Kolodner, R. D. / Renaturation of DNA by a Saccharomyces cerevisiae protein that catalyzes homologous pairing and strand exchange. In: Journal of Biological Chemistry. 1988 ; Vol. 263, No. 29. pp. 15189-15195.
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