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Carcinogenesis Advance Access originally published online on October 29, 2005
Carcinogenesis 2006 27(3):446-453; doi:10.1093/carcin/bgi254
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© The Author 2005. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

An in vivo analysis of MMC-induced DNA damage and its repair

Young-Ju Lee 1, Su-Jung Park 1, 3, Samantha L.M. Ciccone 2, Chong-Rak Kim 4 and Suk-Hee Lee 1, 3, *

1 Department of Biochemistry and Molecular Biology, 2 Microbiology and 3 Walther Oncology Center, Indiana University School of Medicine, Indianapolis, IN 46202, USA and 4 Inje University, Kimhae, Korea

* To whom correspondence should be addressed at: 1044 W. Walnut Street Room 153, Indiana University Cancer Research Institute, Indianapolis, IN 46202, USA. Tel: +1 317 278 3464; Fax: +1 317 274 8046; Email: slee{at}iupui.edu

Mitomycin C (MMC) induces various types of DNA damages that cause significant cytotoxicity to cells. Accordingly, repair of MMC-induced damages involves multiple repair pathways such as nucleotide excision repair, homologous recombination repair and translesion bypass repair pathways. Nonetheless, repair of the MMC-induced DNA damages in mammals have not been fully delineated. In this study, we investigated potential roles for Xeroderma pigmentosum (XP) proteins in the repair of MMC-induced DNA damages using an assay that detects the ssDNA patches generated following treatment with MMC or 8'-methoxy-psoralen (8-MOP) + UVA (ultraviolet light A). Human wild-type cells formed distinctive ssDNA foci following treatment with MMC or 8-MOP + UVA, but not with those inducing alkylation damage, oxidative damage or strand-break damage, suggesting that the foci represent ssDNA patches formed during the crosslink repair. In contrast to wild-type cells, mutant defective in XPE orXPG did not form the ssDNA foci following MMC treatment, while XPF mutant cells showed a significantly delayed response in forming the foci. A positive role for XPG in the repair of MMC-induced DNA damages was further supported by observations that cells treated with MMC induced a tight association of XPG with chromatin, and a targeted inhibition of XPG abolished MMC-induced ssDNA foci formation, rendering cells hypersensitive to MMC. Together, our results suggest that XPG along with XPE and XPF play unique role(s) in the repair of MMC-induced DNA damages.


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