Abstract
Alkylating agents induce genome-wide base damage, which is repaired mainly by N-methylpurine DNA glycosylase (MPG). An elevated expression of MPG in certain types of tumor cells confers higher sensitivity to alkylation agents because MPG-induced apurinic/apyrimidic (AP) sites trigger more strand breaks. However, the determinant of drug sensitivity or insensitivity still remains unclear. Here, we report that the p53 status coordinates with MPG to play a pivotal role in such process. MPG expression is positive in breast, lung and colon cancers (38.7%, 43.4% and 25.3%, respectively) but negative in all adjacent normal tissues. MPG directly binds to the tumor suppressor p53 and represses p53 activity in unstressed cells. The overexpression of MPG reduced, whereas depletion of MPG increased, the expression levels of pro-arrest gene downstream of p53 including p21, 14-3-3σ and Gadd45 but not proapoptotic ones. The N-terminal region of MPG was specifically required for the interaction with the DNA binding domain of p53. Upon DNA alkylation stress, in p53 wild-type tumor cells, p53 dissociated from MPG and induced cell growth arrest. Then, AP sites were repaired efficiently, which led to insensitivity to alkylating agents. By contrast, in p53-mutated cells, the AP sites were repaired with low efficacy. To our knowledge, this is the first direct evidence to show that a DNA repair enzyme functions as a selective regulator of p53, and these findings provide new insights into the functional linkage between MPG and p53 in cancer therapy.
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Acknowledgements
We thank Drs Shengcai Lin (Xiamen University), Bert Vogelstein (Johns Hopkins Oncology Center) and Yue Xiong (University of North Carolina) for providing materials. The study was supported by the National Basic Research Programs (2011CB910802, 2012CB910702, 2010CB912202), and National Natural Science Foundation Projects (31071144, 31125010).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
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Supplementary information, Figure S1
MPG interacts with p53 and inhibits p53 activity in HEK293 cells. (PDF 19 kb)
Supplementary information, Figure S2
p53 has no significant influence on MPG-mediated excision of Hx. (PDF 17 kb)
Supplementary information, Figure S3
MPG selectively downregulates the expression of pro-arrest p53 target genes in HEK293 cells. (PDF 15 kb)
Supplementary information, Data S1
Experimental Protocols (PDF 22 kb)
Supplementary information, Table S1
Primer sequences used for qPCR assays. (PDF 24 kb)
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Song, S., Xing, G., Yuan, L. et al. N-methylpurine DNA glycosylase inhibits p53-mediated cell cycle arrest and coordinates with p53 to determine sensitivity to alkylating agents. Cell Res 22, 1285–1303 (2012). https://doi.org/10.1038/cr.2012.107
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DOI: https://doi.org/10.1038/cr.2012.107
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