title: Nonhomologous end-joining in a cell-free extract from the cultured silkworm cell line BmN4.
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.
authors.affiliation: Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.
authors.forename: Arisa
authors.initials: A
authors.lastname: Ohsaki
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Iiyama
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Miyagawa
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kawaguchi
authors.forename: Katsumi
authors.initials: K
authors.lastname: Koga
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kusakabe
mesh_terms: D000818:Animals; D012831:Bombyx / Q000737:chemistry / Q000166:cytology / Q000235:genetics*; D002413:Cations, Divalent / Q000737:chemistry; D002460:Cell Line; D002474:Cell-Free System; D004249:DNA Damage; D004260:DNA Repair; D008670:Metals / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011995:Recombination, Genetic
chemical_list: D002413:Cations, Divalent; D008670:Metals