Abstract
Programmed cell death (apoptosis) is a conserved process aimed to eliminate unwanted cells. The key molecules are a group of proteases called caspases that cleave vital proteins, which leads to the death of cells. In Drosophila, the apoptotic pathway is usually represented as a cascade of events in which an initial stimulus activates one or more of the proapoptotic genes (hid, rpr, grim), which in turn activate caspases. In stress-induced apoptosis, the dp53 (Drosophila p53) gene and the Jun N-terminal kinase (JNK) pathway function upstream in the activation of the proapoptotic genes. Here we demonstrate that dp53 and JNK also function downstream of proapoptotic genes and the initiator caspase Dronc (Drosophila NEDD2-like caspase) and that they establish a feedback loop that amplifies the initial apoptotic stimulus. This loop plays a critical role in the apoptotic response because in its absence there is a dramatic decrease in the amount of cell death after a pulse of the proapoptotic proteins Hid and Rpr. Thus, our results indicate that stress-induced apoptosis in Drosophila is dependant on an amplification loop mediated by dp53 and JNK. Furthermore, they also demonstrate a mechanism of mutual activation of proapoptotic genes.
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Abbreviations
- Diap1:
-
Drosophila inhibitor of apoptosis protein 1
- dp53 :
-
Drosophila p53
- dp53-i :
-
dp53 RNA interference
- dpp :
-
decapenthaplegic
- Drice:
-
Drosophila interleukin converting enzyme
- Dronc:
-
Drosophila NEDD2-like caspase
- dronc-i:
-
dronc RNA-interference
- en :
-
engrailed
- gfp :
-
green fluorescent protein
- hep :
-
hemipterous
- hid :
-
head involution defective
- hs-hid :
-
heat shock-head involution defective
- hs-rpr :
-
heat shock-reaper
- IAP:
-
inhibitor of apoptosis protein
- JNK:
-
Jun N-terminal kinase
- puc :
-
puckered
- ROI:
-
region of interest
- rpr :
-
reaper
- sal :
-
spalt
- TUNEL:
-
TdT-mediated fluoroscein-dUTP nick end labelling
- UAS:
-
upstream activation sequence
- wg :
-
wingless
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Acknowledgements
We thank Ernesto Sanchez-Herrero for fruitful discussions, Francisco A Martín and Hermann Steller for critical reading of the manuscript and Ana Guarner for help with leg preparations. We also thank Andreas Bergmann for dronc mutants, Hermann Steller for the heat shock stocks, Jose F de Celis for the salEPv-Gal4 line, Don Ryoo for the anti-Hid antibody, Ingrid Lohmann for the rpr probe and the Confocal Microscopy Service at the CBMSO for help with acquisition and processing of images. We also thank Angelica Cantarero and Rosa Gonzalez for general help and the members of the lab for discussions. The experimental work has been funded by grants from the Ministerio de Ciencia e Innovación (Consolider CSD 20700008-B and BFU 2008 03196/BMC) and by an institutional grant from the Fundación Ramón Areces. ES is supported by a FPU scholarship (AP2005-4875).
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Shlevkov, E., Morata, G. A dp53/JNK-dependant feedback amplification loop is essential for the apoptotic response to stress in Drosophila. Cell Death Differ 19, 451–460 (2012). https://doi.org/10.1038/cdd.2011.113
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DOI: https://doi.org/10.1038/cdd.2011.113
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