Abstract
The link between nucleocytoplasmic transport and apoptosis remains controversial. Nucleocytoplasmic exchange of molecules seems indeed essential for the initiation and execution of the apoptotic programme; but inhibition of nuclear transport factors may also represent a powerful apoptotic trigger. The GTPase Ran (together with its partners), first discovered to be essential in nucleocytoplasmic transport, has multiple key functions in cell biology, and particularly in spindle assembly, kinetochore function and nuclear envelope assembly. Among the Ran partners studied, NTF2 appears to be solely involved in nucleocytoplasmic transport. Here, we localised Ran and several of its partners, RanBP1, CAS and NTF2, at the nuclear membrane in the trypanosomatid Leishmania major. Remarkably, these proteins fused to GFP decorated a perinuclear ‘collar’ of about 15 dots, colocalising at nuclear pore complexes with the homologue of nucleoporin Sec13. In the other trypanosomatid Trypanosoma brucei, RNAi knockdown of the expression of the corresponding genes resulted in an apoptosis-like phenomenon. These phenotypes show that Ran and its partners have a key function in trypanosomatids like they have in mammals. Our data, notably those about TbNTF2 RNAi, support the idea that active nucleocytoplasmic transport is not essential for the initiation and execution of apoptosis, and, rather, the impairment of this transport constitutes an intrinsic signal for triggering PCD.
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Abbreviations
- CAS:
-
cellular apoptosis susceptibility protein
- DAPI:
-
4,6-diamidino-2-phenylindole
- FISH:
-
fluorescent in situ hybridisation
- GFP:
-
green fluorescent protein
- NES:
-
nuclear exclusion signal
- NLS:
-
nuclear localisation signal
- NPC:
-
nuclear pore complex
- NTF2:
-
nuclear transport factor 2
- PCD:
-
programmed cell death
- RanBP:
-
Ran-binding protein
- RanGAP:
-
Ran GTPase-activating protein
- RanGEF:
-
Ran guanine exchange factor
- RCC1:
-
regulator of chromosome condensation 1
- RNAi:
-
RNA interference
- TUNEL:
-
terminal uridine deoxynucleotidyl transferase dUTP nick-end labelling
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Acknowledgements
We thank Pierre Travo (Platform RIO Imaging, Montpellier) for expert advice and assistance, as well as Professor Jacques Clot for allowing access to FACS facilities at the Laboratoire d’Immunologie (CHU of Montpellier, France) and François Renaud (UMR2724 CNRS-University Montpellier 1-IRD) for support and encouragement. We also thank Yvon Sterkers notably for the western blots and Laurence Berry-Sterkers for help in microscopy. MC was a recipient of a grant from the Ministère de l’Enseignement Supérieur et de la Recherche.
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Casanova, M., Portalès, P., Blaineau, C. et al. Inhibition of active nuclear transport is an intrinsic trigger of programmed cell death in trypanosomatids. Cell Death Differ 15, 1910–1920 (2008). https://doi.org/10.1038/cdd.2008.132
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DOI: https://doi.org/10.1038/cdd.2008.132
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