Biochemical characterization of endonuclease v from the hyperthermophilic archaeon, Pyrococcus furiosus

Shinichi Kiyonari, Yuriko Egashira, Sonoko Ishino, Yoshizumi Ishino

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18 Citations (Scopus)

Abstract

Endonuclease V (Endo V) is a DNA repair enzyme that recognizes deoxyinosine and cleaves the second phosphodiester bond on the 3′ side of the deaminated base lesion. A database search revealed the presence of homologous genes for Endo V in most archaeal species, but the absence in some methanogenic species. We cloned a gene encoding the sequence homologous to Escherichia coli Endo V from the genome of the hyperthermophilic euryarchaeon, Pyrococcus furiosus and purified gene product (PfuEndoV) to homogeneity. In vitro characterization showed that PfuEndoV possesses specific endonuclease activity for the deoxyinosine-containing DNA strand. The activity of the enzyme was maximal at 90°C. Stable complex formation between PfuEndoV and nicked DNA produced by the cleavage reaction was detected by gel mobility shift assays. The molecular mechanisms of the inosine repair pathway including Endo V in the archaeal cells are discussed. Interestingly, PfuEndoV cleaved inosine-containing RNA strands as well as DNA substrates. PfuEndoV may also be involved in RNA metabolism.

Original languageEnglish
Pages (from-to)325-333
Number of pages9
JournalJournal of biochemistry
Volume155
Issue number5
DOIs
Publication statusPublished - May 2014

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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