In situ analysis of repair processes for oxidative DNA damage in mammalian cells

Li Lan, Satoshi Nakajima, Yoshitsugu Oohata, Masashi Takao, Satoshi Okano, Mitsuko Masutani, Samuel H. Wilson, Akira Yasui

Research output: Contribution to journalArticlepeer-review

227 Citations (Scopus)

Abstract

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 β 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 polymerase β-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.

Original languageEnglish
Pages (from-to)13738-13743
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number38
DOIs
Publication statusPublished - 2004 Sep 21

ASJC Scopus subject areas

  • General

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