Abstract
Previous biochemical studies suggested that HIV-1-encoded Vpr may kill cells through an effect on the adenine nucleotide translocase (ANT), thereby causing mitochondrial membrane permeabilization (MMP). Here, we show that Vpr fails to activate caspases in conditions in which it induces cell killing. The knock-out of essential caspase-activators (Apaf-1 or caspase-9) or the knock-out of a mitochondrial caspase-independent death effector (AIF) does not abolish Vpr-mediated killing. In contrast, the cytotoxic effects of Vpr are reduced by transfection-enforced overexpression of two MMP-inhibitors, namely the endogenous protein Bcl-2 or the cytomegalovirus-encoded ANT-targeted protein vMIA. Vpr, which can elicit MMP through a direct effect on mitochondria, and HIV-1-Env, which causes MMP through an indirect pathway, exhibit additive (but not synergic) cytotoxic effects. In conclusion, it appears that Vpr induces apoptosis through a caspase-independent mitochondrial pathway.
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Abbreviations
- AIF:
-
apoptosis-inducing factor
- ANT:
-
adenine nucleotide translocator
- COX:
-
cytochrome c oxidase
- (Asp)2R110:
-
bis-(L-aspartic acid amide) rhodamine 110
- CMV:
-
cytomegalovirus
- Cyt c:
-
cytochrome c
- DiOC6(3):
-
3,3′dihexyloxacarbocyanine iodide
- ΔΨm:
-
mitochondral transmembrane potential
- ES:
-
embryonic stem
- JC-1:
-
5,5′,6,6′-tetrachloro-1,1′, 3,3′-tetraethylbenzimidazolylcarbocyanine iodide
- MMP:
-
mitochondrial membrane permeabilization
- PI:
-
propidium iodide
- SIMP:
-
soluble intermembrane protein
- STS:
-
staurosporin
- VDAC:
-
voltage-dependent anion channel
- vMIA:
-
viral mitochondrial inhibitor of apoptosis
- Z-VAD.fmk:
-
N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone
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Acknowledgements
We thank Dr. Victor Goldmacher (Apoptosis Technology Inc., Cambridge, MA, USA) for vMIA and Bcl-2-transfected HeLa cells, Dr. Tak Mak (The Amgen and Ontario Cancer Institute, Toronto, Canada) for Apaf-1−/− and caspase-9−/− cells, Etienne Jacotot for advice, and Didier Métivier (Villejuif, France) for technical assistance. This work has been supported by a special grant from the Ligue Nationale contre le Cancer, as well as grants from ANRS, Sidaction and the European Commission (QLG1-CT-1999-00739 to G Kroemer). HLA Vieira receives a fellowship from the Fundação para a Ciência e a Tecnologia PRAXIS XXI, Portugal; P Boya from the European Commission (MCFI-2000-00943); KF Ferri from MRT.
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Roumier, T., Vieira, H., Castedo, M. et al. The C-terminal moiety of HIV-1 Vpr induces cell death via a caspase-independent mitochondrial pathway. Cell Death Differ 9, 1212–1219 (2002). https://doi.org/10.1038/sj.cdd.4401089
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DOI: https://doi.org/10.1038/sj.cdd.4401089
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