Abstract
In mammals and Drosophila, apoptotic caspases are under positive control of the CED-4-like proteins Apaf-1 and ARK, respectively. In an EMS-mutagenesis screen, we isolated 33 ark mutants as recessive suppressors of hid-induced apoptosis. The ark mutants are loss-of-function alleles characterized by reduced developmental apoptosis. Using the phenotypic series of these alleles, we identified helical domain I in the nucleotide oligomerization domain as critical for ARK's apoptotic activity. Interestingly, the WD40 region may also have an unanticipated positive requirement for the apoptotic activity of ARK. Considering structural information, we discuss the roles of these domains for assembly and activity of the ARK apoptosome, and propose that the WD40 region is anti-apoptotic in the absence of apoptotic signals, and pro-apoptotic in the presence of such signals. Furthermore, a defined null allele reveals that ark is required for most, but not all apoptosis suggesting the existence of an ARK-independent apoptotic pathway.
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Abbreviations
- aa:
-
amino acid
- AAA+:
-
ATPases associated with various cellular activities
- AO:
-
acridine orange
- Apaf-1:
-
apoptotic protease activating factor-1
- ARK:
-
Apaf-1 related killer
- BrdU:
-
bromodeoxyuridine
- CARD:
-
caspase activation and recruitment domain
- CED-4:
-
cell death deficient-4
- CTD:
-
C-terminal domain
- DFS:
-
dominant female sterile
- DIAP1:
-
Drosophila inhibitor of apoptosis protein 1
- DREDD:
-
death-related ced-3/Nedd2-like protein
- DrICE:
-
Drosophila ICE
- DRONC:
-
Drosophila Nedd-2 like Caspase
- EMS:
-
ethyl-methyl sulfonate
- ey :
-
eyeless
- FLP:
-
Flippase
- FRT:
-
Flippase recombination target
- GheF:
-
GMR-hid ey-Flp
- GLC:
-
germline clones
- GMR:
-
glass multimer reporter
- HD1:
-
helix domain 1
- HD2:
-
helix domain 2
- hid :
-
head involution defective
- IAP:
-
inhibitor of apoptosis proteins
- kDa:
-
kilo Dalton
- MG:
-
midline glia
- MF:
-
morphogenetic furrow
- NMD:
-
non-sense mRNA-mediated decay
- NOD:
-
nucleotide oligomerization domain
- RHG:
-
Reaper Hid Grim
- SMW:
-
second mitotic wave
- TUNEL:
-
terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling
- wg:
-
wingless
- WHD:
-
winged-helix domain
- ZCP:
-
zone of proliferating cells
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Acknowledgements
We apologize to all our colleagues whose work could not be cited due to space constraints. We thank George Jackson for the anti-ARK antibody; John Abrams for the arkCD4 allele and sharing information before publication; Hyung-Don Ryoo and Hermann Steller for antibodies; Bruce Hay and the Bloomington stock center for fly stocks; Eli Arama for pointing out that arkL46/arkP46 mutants produce viable adults; the MD Anderson DNA Analysis Core Facility for sequencing of ark alleles (supported by Core Grant No. CA16672 from the NCI). AB is a fellow of the MD Anderson Research Trust. This work was supported by grants from the NIH (GM068016) and The Robert A. Welch Foundation (G-1496) to AB.
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Srivastava, M., Scherr, H., Lackey, M. et al. ARK, the Apaf-1 related killer in Drosophila, requires diverse domains for its apoptotic activity. Cell Death Differ 14, 92–102 (2007). https://doi.org/10.1038/sj.cdd.4401931
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DOI: https://doi.org/10.1038/sj.cdd.4401931
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