Abstract
The p53 tumor suppressor responds to certain cellular stresses by inducing transcriptional programs that can lead to growth arrest or apoptosis. However, the molecular mechanisms responsible for choosing between these two cell fates are not well understood. Previous studies have suggested that p53 selectively activates proarrest target genes, due to the higher affinity of p53 for their promoters compared with proapoptotic genes. Here we show using microarray and chromatin immunoprecipitation that p53 binds to and transcriptionally activates both its proarrest and proapoptotic target genes proportionally to induced p53 expression levels. Further, we provide evidence that to trigger apoptosis, cells must overcome an apoptotic threshold, whose height is determined by expression levels of p53 and its targets, the duration of their expression and the cellular context. We demonstrate in multiple cells lines that below this threshold, expression levels of p53 and its targets were sufficient to induce arrest but not apoptosis. Above this threshold, p53 and its targets triggered extensive apoptosis. Moreover, lowering this threshold with inhibitors of antiapoptotic Bcl-2 family proteins sensitized cells to p53-induced apoptosis. These findings argue that agents that lower the apoptotic threshold should increase the efficacy of p53-mediated cancer therapy.
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Accession codes
Abbreviations
- ChIP:
-
chromatin immunoprecipitation
- dox:
-
doxycycline
- HMEC:
-
human mammary epithelial cells
- γH2AX:
-
phosphorylated form of H2AX
- PI:
-
propidium iodide
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Acknowledgements
We are grateful to Dr. Bo Zhao for his help with cloning procedures. Confocal laser scanning microscopy was performed at the Mount Sinai School of Medicine Microscopy Shared Resources Facility, supported with funding from NIH-NCI shared resources (Grant no. 5R24CA095823-04), National Science Foundation Major Research Instrumentation (Grant no. DBI-9724504) and NIH shared instrumentation (Grant no. 1 S10RR0 9145-01). We are also grateful to Dr. Huei-Chi Wen for his help with the confocal imaging analysis. We thank Luis Carvajal and Dr. Pierre-Jacques Hamard from Dr. James J Manfredi’s lab for sharing their ChIP assay expertise and providing us with the ChIP MDM2, p21 and PUMA primer sequences, Dr. Greg Khitrov in the Departmental Core Life Sciences Technology Laboratory of the Mount Sinai School of Medicine for performing the hybridization and processing of our Gene 1.1 ST Arrays, and Dr. Sam W Lee for helpful discussion. This study was supported by grants from the Breast Cancer Research Foundation, the Empire State Stem Cell Fund through New York State Department of Health Contract no. C024313 (SAA) and by P01 CA080058 from the National Cancer Institute (SAA). The content is solely the responsibility of the authors and does not necessarily represent the official views of the Empire State Stem Cell Board, the New York State Department of Health, the State of New York, the National Cancer Institute or the National Institutes of Health. Supplementary Information is available at Cell Death and Differentiation’s website.
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Kracikova, M., Akiri, G., George, A. et al. A threshold mechanism mediates p53 cell fate decision between growth arrest and apoptosis. Cell Death Differ 20, 576–588 (2013). https://doi.org/10.1038/cdd.2012.155
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DOI: https://doi.org/10.1038/cdd.2012.155
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