TY - JOUR
T1 - Nonhomologous end-joining in a cell-free extract from the cultured silkworm cell line BmN4.
AU - Ohsaki, Arisa
AU - Iiyama, Kazuhiro
AU - Miyagawa, Yoshitaka
AU - Kawaguchi, Yutaka
AU - Koga, Katsumi
AU - Kusakabe, Takahiro
N1 - Funding Information:
This work was supported in part by grants (No. 14704010 and No. 1456024) and National Biore-source Project from the Ministry of Education, Science and Culture of Japan.
PY - 2005/3
Y1 - 2005/3
N2 - 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.
AB - 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.
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U2 - 10.1007/s11033-004-2474-y
DO - 10.1007/s11033-004-2474-y
M3 - Article
C2 - 15865207
AN - SCOPUS:24744444368
VL - 32
SP - 25
EP - 34
JO - Molecular Biology Reports
JF - Molecular Biology Reports
SN - 0301-4851
IS - 1
ER -