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  • Title: Identification and characterization of a mitochondrial endonuclease from yeast, Schizosaccharomyces pombe.
    Author: Ikeda S, Maeda N, Ohshima T, Takata N.
    Journal: Biochem Mol Biol Int; 1996 Nov; 40(5):1017-24. PubMed ID: 8955892.
    Abstract:
    Schizosaccharomyces pombe mitochondria were isolated from the cells treated with Novozyme 234, and purified in a Percoll gradient. A zymographic assay in a SDS-polyacrylamide gel containing single-stranded DNA revealed that an endonuclease of 32 kDa is associated with the mitochondria. The endonuclease was extracted from the mitochondria with 0.5 M KCl and was partially purified. The 32-kDa enzyme degraded both DNA and RNA at a weak alkaline pH, but preferred single-stranded DNA. The enzyme required Mg2+ or Mn2+, but not Ca2+ or Zn2+ for activity, and was inhibited by 50% with a 150 mM salt solution. Nicks generated by the enzyme could be resealed with T4 DNA ligase, indicating that the enzyme produces 5'-P and 3'-OH ends.
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