A multifunctional exonuclease from vegetative schizosaccharomyces pombe cells exhibiting in vitro strand exchange activity

Edgar Käslin, Wolf Dietrich Heyer

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

A 140-kDa polypeptide (p140) has been purified over 2000-fold from vegetative Schizosaccharomyces pombe cells using an assay of homologous pairing and strand exchange between linear double-stranded DNA (dsDNA) and circular single-stranded DNA (ssDNA) in vitro. Electron microscopic analysis of the reaction products showed displacement of one strand of the linear duplex DNA by the circular ssDNA molecule. In addition, the protein contained 5' to 3' exonuclease activity on ssDNA and dsDNA (with a 50-fold preference on the single-stranded substrate) as well as on single-stranded RNA. Furthermore, p140 was capable of renaturing complementary ssDNA as shown by S1 nuclease assays. p140 behaved like a monomer in solution under reaction conditions. Direct comparison of the biochemical properties, sequence analysis, and cross-reactivity to a monoclonal antibody suggests that p140 is probably identical with Exoll, purified from S. pombe meiotic cells as a ssDNA exonuclease (Szankasi, P., and Smith, G. R. (1992) Biochemistry 31, 6769-6773). Given the diverse activities of p140, the protein might be involved in DNA and/or RNA metabolism in vivo.

Original languageEnglish (US)
Pages (from-to)14094-14102
Number of pages9
JournalJournal of Biological Chemistry
Volume269
Issue number19
StatePublished - May 13 1994
Externally publishedYes

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Exonucleases
Schizosaccharomyces
Single-Stranded DNA
spleen exonuclease
Assays
Exodeoxyribonucleases
RNA
Circular DNA
Biochemistry
Reaction products
Metabolism
Sequence Analysis
Proteins
Complementary DNA
Monomers
Monoclonal Antibodies
Electrons
Peptides
Molecules
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

A multifunctional exonuclease from vegetative schizosaccharomyces pombe cells exhibiting in vitro strand exchange activity. / Käslin, Edgar; Heyer, Wolf Dietrich.

In: Journal of Biological Chemistry, Vol. 269, No. 19, 13.05.1994, p. 14094-14102.

Research output: Contribution to journalArticle

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