Abstract
The ability of p53 to induce apoptosis in cells with damaged DNA is thought to contribute greatly to its tumour suppressor function. P53 has been proposed to induce apoptosis via numerous transcriptional targets or even by direct cytoplasmic action. Two transcriptional targets shown to mediate its apoptotic role in several cell types encode Noxa and Puma, BH3-only members of the Bcl-2 family. To test if their functions in p53-dependent apoptosis overlap, we generated mice lacking both. These mice develop normally and no tumours have yet arisen. In embryonic fibroblasts, the absence of both Noxa and Puma prevented induction of apoptosis by etoposide. Moreover, following whole body γ-irradiation, the loss of both proteins protected thymocytes better than loss of Puma alone. Indeed, their combined deficiency protected thymocytes as strongly as loss of p53 itself. These results indicate that, at least in fibroblasts and thymocytes, p53-induced apoptosis proceeds principally via Noxa and Puma, with Puma having the predominant role in diverse cell types. The absence of tumours in the mice suggests that tumour suppression by p53 requires functions in addition to induction of apoptosis.
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Abbreviations
- BH:
-
Bcl-2-homology
- DKO:
-
double knockout
- DP:
-
double-positive
- MEF:
-
mouse embryonic fibroblasts
- PMA:
-
phorbol 12-myristate 13-acetate
- TUNEL:
-
terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling
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Acknowledgements
We thank N Iannarella, G Siciliano and A Naughton for animal care; Dr. F Battye, C Tarlinton, V Milovac, J Garbe and C Young for cell sorting; T Nikolaou and G Thomas for γ-irradiation; J Corbin for haematological analysis; Dr. S Mihajlovic, A Hasanein, K Weston for histological sections; Drs. M Schuler, S Lowe and G Hannon for expression vectors; and Professor S Cory and Drs. A Harris, D Huang, H Puthalakath, P Bouillet, L O’Reilly, M Pellegrini, L Coultas and C Scott for their input and interesting discussions. This work was supported by fellowships and grants from the NHMRC (Canberra; program #257502), the Leukemia and Lymphoma Society (SCOR grant #7015), the NIH (CA043540-18 and CA80188-6), the JDRF/NHMRC, the Leukemia Research Foundation (LRF), the Cancer Council Victoria Postdoctoral Cancer Research Fellowship and the Austrian Science Fund (FWF).
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Michalak, E., Villunger, A., Adams, J. et al. In several cell types tumour suppressor p53 induces apoptosis largely via Puma but Noxa can contribute. Cell Death Differ 15, 1019–1029 (2008). https://doi.org/10.1038/cdd.2008.16
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DOI: https://doi.org/10.1038/cdd.2008.16
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