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In situ analysis of repair processes for oxidative DNA damage in mammalian cells
被引:248
作者:
Lan, L
Nakajima, S
Oohata, Y
Takao, M
Okano, S
Masutani, M
Wilson, SH
Yasui, A
机构:
[1] Tohoku Univ, Dept Mol Genet, Inst Dev Aging & Canc, Sendai, Miyagi 9808575, Japan
[2] Yamagata Univ, Res Lab Mol Genet, Yamagata 9909585, Japan
[3] Natl Canc Ctr, Res Inst, Div Biochem, Tokyo 1040045, Japan
[4] NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA
来源:
关键词:
D O I:
10.1073/pnas.0406048101
中图分类号:
O [数理科学和化学];
P [天文学、地球科学];
Q [生物科学];
N [自然科学总论];
学科分类号:
07 ;
0710 ;
09 ;
摘要:
Oxidative DNA damage causes blocks and errors in transcription and replication, leading to cell death and genomic instability. Although repair mechanisms of the damage have been extensively analyzed in vitro, the actual in vivo repair processes remain largely unknown. Here, by irradiation with an UVA laser through a microscope lens, we have conditionally produced single-strand breaks and oxidative base damage at restricted nuclear regions of mammalian cells. We showed, in real time after irradiation by using antibodies and GFP-tagged proteins, rapid and ordered DNA repair processes of oxidative DNA damage in human cells. Furthermore, we characterized repair pathways by using repair-defective mammalian cells and found that DNA polymerase 13 accumulated at single-strand breaks and oxidative base damage by means of its 31- and 8-kDa domains, respectively, and that XRCC1 is essential for both polymerarse beta-dependent and proliferating cell nuclear antigen-dependent repair pathways of single-strand breaks. Thus, the repair of oxidative DNA damage is based on temporal and functional interactions among various proteins operating at the site of DNA damage in living cells.
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页码:13738 / 13743
页数:6
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