Abstract
Nitric oxide (NO) is a potent activator of the p53 tumor suppressor protein. However, the mechanisms underlying p53 activation by NO have not been fully elucidated. We previously reported that a rapid downregulation of Mdm2 by NO may contribute to the early phase of p53 activation. Here we show that NO promotes p53 nuclear retention and inhibits Mdm2-mediated p53 nuclear export. NO induces phosphorylation of p53 on serine 15, which does not require ATM but rather appears to depend on the ATM-related ATR kinase. An ATR-kinase dead mutant or caffeine, which blocks the kinase activity of ATR, effectively abolishes the ability of NO to cause p53 nuclear retention, concomitant with its inhibition of p53 serine 15 phosphorylation. Of note, NO enhances markedly the ability of low-dose ionizing radiation to elicit apoptotic killing of neuroblastoma cells expressing cytoplasmic wild-type p53. These findings imply that, through augmenting p53 nuclear retention, NO can sensitize tumor cells to p53-dependent apoptosis. Thus, NO donors may potentially increase the efficacy of radiotherapy for treatment of certain types of cancer.
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Abbreviations
- NO:
-
nitric oxide
- NOS:
-
nitric oxide synthases
- NES:
-
nuclear export signal
- PARP:
-
Poly(ADP-Ribose) Polymerase 1
- NLS:
-
nuclear localization signals
- LMB:
-
lyptomycin B
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Acknowledgements
We thank Dr. D Michael for helpful discussions, Dr. Y Taya for the anti-Ser15-P antibody, Dr. M Schwab and Dr. Y Shiloh and for IMR32 and ap24 cells, respectively, Dr. M Kastan for pCDNA3-ATR-wt and pCDNA3-ATR-kd plasmids and Dr. B Geiger for anti-vinculin antibodies. This work was supported in part by grant RO1 CA 40099 from the National Cancer Institute (USA), The USA–Israel Binational Science Foundation, the Kadoorie Charitable Foundations, the Center for Excellence Program of the Israel Science Foundation, and the Yad Abraham Center for Cancer Diagnosis and Therapy.
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Wang, X., Zalcenstein, A. & Oren, M. Nitric oxide promotes p53 nuclear retention and sensitizes neuroblastoma cells to apoptosis by ionizing radiation. Cell Death Differ 10, 468–476 (2003). https://doi.org/10.1038/sj.cdd.4401181
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DOI: https://doi.org/10.1038/sj.cdd.4401181
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