Abstract
PUMA, a key mediator of p53-induced apoptosis, is a BH3-only domain proapoptotic protein that localizes to mitochondria and interacts with antiapoptotic Bcl-2 and Bcl-XL. Recent evidence implicates Bax to be an important mediator of PUMA-activated apoptotic signals. We have previously demonstrated that Bax deficiency significantly affects thapsigargin (TG)-mediated endoplasmic reticulum calcium pool depletion-induced apoptosis. We now present evidence that TG upregulates PUMA expression and that although Bax-deficient cells exhibit resistance to TG, Bax deficiency does not attenuate TG upregulation of PUMA expression. Furthermore, TG transcriptionally upregulates PUMA expression in a p53-independent manner and that PUMA-deficient cells are more resistant to undergo TG-induced apoptosis than the PUMA-proficient counterparts. Thus, our results demonstrate that TG engages PUMA and Bax for full transduction of apoptotic signals and both PUMA and Bax appear to exist in the same TG-activated apoptotic pathway in which PUMA may reside upstream of Bax.
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Abbreviations
- Ca2+:
-
calcium
- CaM:
-
calmodulin
- CREB:
-
cAMP response element-binding protein
- bbc3:
-
Bcl-2 binding component 3
- DMEM:
-
Dulbecco's modified Eagle's medium
- DKO:
-
double knockout
- DR5:
-
death receptor 5
- ER:
-
endoplasmic reticulum
- ERSEs:
-
ER stress response elements
- ETOP:
-
etoposide
- InsP3R:
-
inositol (1,4,5) triphosphate receptor
- PBS:
-
phospate-buffered saline
- grp:
-
glucose-regulated protein
- PUMA:
-
p53 upregulated modulator of apoptosis
- TG:
-
thapsigargin
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Acknowledgements
We thank Dr. Bert Vogelstein (Johns Hopkins University, Baltimore, MD, USA) for kindly providing the Bax and PUMA-proficient and -deficient HCT116 cells, and Dr. Allen (SUNY Upstate Medical University, Syracuse, NY, USA) and Dr. Oscar Colamonici (The University of Illinios at Chicago, Chicago, IL, USA) for Saos-2 cells. We are also thankful to Dr. Chittenden (ImmunoGen, Cambridge, MA, USA) for providing the human PUMA/bbc promoter-luciferase construct, and Dr. Richard J Youle (National Institutes of Health, Bethesda, MD, USA) for providing the pEGFP-Bax construct. This work was partly supported by the NIH Grants CA86945, DK062136, DK067271 and funds from the Sinsheimer Scholar Award by the Alxandrine and Alexander Sinsheimer Foundation.
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Luo, X., He, Q., Huang, Y. et al. Transcriptional upregulation of PUMA modulates endoplasmic reticulum calcium pool depletion-induced apoptosis via Bax activation. Cell Death Differ 12, 1310–1318 (2005). https://doi.org/10.1038/sj.cdd.4401659
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DOI: https://doi.org/10.1038/sj.cdd.4401659
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