Abstract
Previous studies by our laboratory have shown that the drug transporter protein P-glycoprotein, P-gp, can specifically inhibit Fas-induced caspase-3 activation and apoptosis. Importantly, inhibition of both caspase-3 activation and cell death could be reversed by pharmacological and antibody inhibitors of P-gp function. However, the molecular mechanisms underpinning P-gp-mediated resistance to Fas-induced cell death and caspase activation remained unknown. We therefore sought to identify the point(s) within the death receptor pathway at which P-gp exerted its inhibitory effect and to determine whether the ATPase activity of P-gp was required. Structure-function analysis determined that ATP hydrolysis was necessary for P-gp to confer resistance to Fas-induced caspase activation and cell death. Importantly, although both FADD and caspase-8 were recruited to the Death Inducing Signal Complex (DISC) in wild-type P-gp expressing cells following Fas ligation, subsequent activation of caspase-8 at the DISC was inhibited. The ability of P-gp to inhibit caspase-8 activation was also ATP dependent. These studies demonstrate that P-gp inhibits Fas-induced caspase-8 activation but not formation of the DISC and that this activity of P-gp is dependent on ATP hydrolysis.
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Abbreviations
- P-gp:
-
P-glycoprotein
- DISC:
-
death inducing signaling complex
- MDR:
-
multidrug resistance
- TNF:
-
tumor necrosis factor
- GFP:
-
green fluorescence protein
- FLIP:
-
Flice-like inhibitory protein
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Acknowledgements
The authors thank Igor Roninson, Paul Roepe and Steve Jane for providing reagents, Ralph Rossi for technical support, and Sarah Russell and Joe Trapani for helpful discussions. RW Johnstone is a Wellcome Trust Senior Research Fellow and MJ Smyth is a Principal Research Fellow of the National Health and Medical Research Council of Australia. This work is supported by a project grant from the NH&MRC, the Anti-Cancer Council of Victoria (ACCV) and the Wellcome Trust.
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Ruefli, A., Tainton, K., Darcy, P. et al. P-glycoprotein inhibits caspase-8 activation but not formation of the death inducing signal complex (disc) following Fas ligation. Cell Death Differ 9, 1266–1272 (2002). https://doi.org/10.1038/sj.cdd.4401081
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DOI: https://doi.org/10.1038/sj.cdd.4401081
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