Abstract
Cell division cycle 25A (Cdc25A) was shown to colocalise both with nuclear and cytoplasmic proteins. Recently, we have demonstrated that overexpressed Cdc25A promoted the survival of rat 423 cells through indirect activation of PKB-protein kinase B. Using a Cdc25A:ER fusion protein, which can be shuttled from the cytoplasm into the nucleus, the present investigation evidences that the antiapoptotic effect of Cdc25A was restricted to its cytoplasmic localisation in rat 423 cells. In contrast, nuclear Cdc25A overexpression caused dephosphorylation and nuclear retention of the proapoptotic transcription factor Forkhead in rhabdomyosarcoma-like 1 (FKHRL1) in human N.1 ovarian carcinoma cells. This resulted in the increased constitutive expression of the FKHRL1 targets Fas ligand and Bim, and promoted apoptosis. Thus, the Cdc25A oncogene, which was found to be frequently overexpressed in certain human cancers, can increase or decrease the susceptibility to apoptosis depending on the cell-type-specific subcellular distribution.
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Abbreviations
- Akt:
-
PKB-protein kinase B
- ASK:
-
apoptosis signal-regulating kinase
- ATP:
-
adenosine triphosphate
- Bim:
-
proapoptotic member of the Bcl-2 family of proteins
- Cdc25A:
-
cell division cycle 25A
- Cdk2:
-
cyclin-dependent kinase 2
- DAPI:
-
4′-6-diamino-2-phenylindol
- DTT:
-
DL-dithiothreitol
- EDTA:
-
ethylenediaminetetraacetic acid
- ER:
-
oestrogen receptor
- FasL:
-
Fas ligand
- FCS:
-
foetal calf serum
- FITC:
-
fluorescein isothiocyanate
- GST:
-
glutathione-S-transferase
- FKHRL1:
-
Forkhead in rhabdomyosarcoma-like 1
- HEPES:
-
(N-[2-hydroxyethyl]piperazine-N′-[2-ethanesulphonic acid])
- HO:
-
Hoechst 33258
- 4-OHT:
-
4-hydroxytamoxifen
- IPTG:
-
(isopropyl[beta]-D-thiogalactoside)
- NLS:
-
nuclear localisation signal
- PBS:
-
phosphate-buffered saline
- PI:
-
propidium iodide
- PMSF:
-
phenylmethylsulphonyl fluoride
- PTEN:
-
phosphatase dephosphorylating phosphatidylinositol 3,4,5-trisphosphate (PIP3)
- SDS:
-
sodium dodecyl sulphate
- TNFa:
-
tumour necrosis factor alpha
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Acknowledgements
We thank the donators providing plasmids and cell lines (listed under ‘Materials and Methods’) and Anton Jaeger for helping with the figures. This work was supported in part by Jubilaeumsfonds der Oesterreichischen Nationalbank, Grant No. 9298, the Herzfelder'sche Familienstiftung and by the Unruhe Privatstiftung (to GK), by the Theodor Koerner Stiftungspreis (to GF) and by a research fellowship of the Medical Faculty of the University of Vienna (to MG).
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Leisser, C., Rosenberger, G., Maier, S. et al. Subcellular localisation of Cdc25A determines cell fate. Cell Death Differ 11, 80–89 (2004). https://doi.org/10.1038/sj.cdd.4401318
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DOI: https://doi.org/10.1038/sj.cdd.4401318
