Apoptosis

, Volume 15, Issue 12, pp 1425–1434 | Cite as

Drosophila Chk2 and p53 proteins induce stage-specific cell death independently during oogenesis

  • Anna Bakhrat
  • Tracy Pritchett
  • Gabriella Peretz
  • Kimberly McCall
  • Uri Abdu
Original Paper

Abstract

In Drosophila, the checkpoint protein-2 kinase (DmChk2) and its downstream effector protein, Dmp53, are required for DNA damage-mediated cell cycle arrest, DNA repair and apoptosis. In this study we focus on understanding the function of these two apoptosis inducing factors during ovarian development. We found that expression of Dmp53, but not DmChk2, led to loss of ovarian stem cells. We demonstrate that expression of DmChk2, but not Dmp53, induced mid-oogenesis cell death. DmChk2 induced cell death was not suppressed by Dmp53 mutant, revealing for the first time that in Drosophila, over-expression of DmChk2 can induce cell death which is independent of Dmp53. We found that over-expression of caspase inhibitors such as DIAP1, p35 and p49 did not suppress DmChk2- and Dmp53-induced cell death. Thus, our study reveals stage-specific effects of Dmp53 and DmChk2 in oogenesis. Moreover, our results demonstrate that although DmChk2 and Dmp53 affect different stages of ovarian development, loss of ovarian stem cells by p53 expression and mid-oogenesis cell death induced by DmChk2 do not require caspase activity.

Keywords

Drosophila melanogaster Ovary Chk2 p53 Caspase Cell death 

Notes

Acknowledgments

We thank Trudi Schüpbach, Andreas Bergmann, Thomas Neufeld, Michael Brodsky and the Bloomington stock center for generously providing fly strains and reagents. This research was supported by Israel Cancer association grant (to UA) and NIH R01 GM60574 (to KM).

Supplementary material

10495_2010_539_MOESM1_ESM.tif (1.5 mb)
Fig. S1DmChk2 genetically interacts with Dmp53 in regulating apoptosis in the Drosophila eye. ad Scanning electron micrographs of eyes from GMR-Gal4 flies crossed to a wild-type fly (a), UASp-DmChk2 fly (b), UASp-Dmp53 fly (c), or UASp-Dmp53 UASp-DmChk2 fly (d). Expression of DmChk2 in the Drosophila eye had no effect on eye morphology (Fig. 1b). We found that overexpression of Dmp53 in the eye using GMR-Gal4 result in a reduced eye size with partial fusion of the ommatidia and some remaining bristles due to apoptosis (Fig. 1c). Co-expression of DmChk2 and Dmp53 resulted in a considerably more severe phenotype with almost complete loss of the eye compared to flies expressing Dmp53 alone (Fig. 1d). We repeated these experiments using all of our transgenic flies and found similar phenotypes. (TIFF 1585 kb)

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Anna Bakhrat
    • 1
    • 3
  • Tracy Pritchett
    • 2
  • Gabriella Peretz
    • 1
    • 3
  • Kimberly McCall
    • 2
  • Uri Abdu
    • 1
    • 3
  1. 1.Department of Life SciencesBen-Gurion UniversityBeer-ShevaIsrael
  2. 2.Department of BiologyBoston UniversityBostonUSA
  3. 3.National Institute for Biotechnology in the NegevBen-Gurion UniversityBeer-ShevaIsrael

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