Nonhomologous end-joining in a cell-free extract from the cultured silkworm cell line BmN4.

Arisa Ohsaki, Kazuhiro Iiyama, Yoshitaka Miyagawa, Yutaka Kawaguchi, Katsumi Koga, Takahiro Kusakabe

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Nonhomologous end-joining (NHEJ) is one of the repair pathways for double-strand breaks (DSBs) in eukaryotic cells. By using linearized plasmid substrates, we have detected intramolecular NHEJ activity in a cell-free extract from the cultured silkworm cell line BmN4. The efficiency of NHEJ differed according to the structure of DNA ends; approximately 1% of input DNA was repaired when the substrate had cohesive ends. The reaction required the hydrolysis of nucleotide triphosphate; interestingly, all of four rNTPs or four dNTPs could support the reaction. A substrate with non-complementary DNA ends was mainly repaired by the DNA polymerase-mediated pathway. These results indicate that the present cell-free system will be useful to analyze the molecular mechanisms of DSB repair and NHEJ in insect cells.

Original languageEnglish
Pages (from-to)25-34
Number of pages10
JournalMolecular biology reports
Volume32
Issue number1
DOIs
Publication statusPublished - Jan 1 2005

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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