Apoptosome-Independent Activation of the Lysosomal Cell Death Pathway by Caspase-9 ^,

Mads Gyrd-Hansen,^1,2, Thomas Farkas,¹ Nicole Fehrenbacher,¹ Lone Bastholm,² Maria Høyer-Hansen,¹ Folmer Elling,² David Wallach,³ Richard Flavell,⁴ Guido Kroemer,⁵ Jesper Nylandsted,¹, and Marja Jäättelä^1*

Apoptosis Department and Centre for Genotoxic Stress, Institute for Cancer Biology, Danish Cancer Society, DK-2100 Copenhagen, Denmark,¹ Institute of Molecular Pathology, University of Copenhagen, DK-2100 Copenhagen, Denmark,² Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel,³ Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, Connecticut 06520,⁴ Centre National de la Recherche Scientifique, UMR 8125, Institut Gustave Roussy, F-94805 Villejuif, France⁵

Abstract: The apoptosome, a heptameric complex of Apaf-1, cytochrome c, and caspase-9, has been considered indispensable for the activation of caspase-9 during apoptosis. By using a large panel of genetically modified murine embryonic fibroblasts, we show here that, in response to tumor necrosis factor (TNF), caspase-8 cleaves and activates caspase-9 in an apoptosome-independent manner. Interestingly, caspase-8-cleaved caspase-9 induced lysosomal membrane permeabilization but failed to activate the effector caspases whereas apoptosome-dependent activation of caspase-9 could trigger both events. Consistent with the ability of TNF to activate the intrinsic apoptosis pathway and the caspase-9-dependent lysosomal cell death pathway in parallel, their individual inhibition conferred only a modest delay in TNF-induced cell death whereas simultaneous inhibition of both pathways was required to achieve protection comparable to that observed in caspase-9-deficient cells. Taken together, the findings indicate that caspase-9 plays a dual role in cell death signaling, as an activator of effector caspases and lysosomal membrane permeabilization.

* Corresponding author. Mailing address: Apoptosis Department and Centre for Genotoxic Stress, Institute for Cancer Biology, Danish Cancer Society, Strandboulevarden 49, DK-2100 Copenhagen, Denmark. Phone: 45-35257318. Fax: 45-35257721. E-mail: mj{at}cancer.dk.

Supplemental material for this article may be found at http://mcb.asm.org/.

Present address: The Breakthrough Toby Robins Breast Cancer Research Centre at The Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, United Kingdom.

The Endolysosomal System in Cell Death and Survival.

U. Repnik, M. H. Cesen, and B. Turk (2013)
Cold Spring Harb Perspect Biol 5, a008755
 |  Abstract »  |  Full Text »  |  PDF »

A variable cytoplasmic domain segment is necessary for {gamma}-protocadherin trafficking and tubulation in the endosome/lysosome pathway.

R. O'Leary, J. E. Reilly, H. H. Hanson, S. Kang, N. Lou, and G. R. Phillips (2011)
Mol. Biol. Cell 22, 4362-4372
 |  Abstract »  |  Full Text »  |  PDF »

Lysosomal-Mitochondrial Axis in Zoledronic Acid-induced Apoptosis in Human Follicular Lymphoma Cells.

L. M. Mitrofan, F. B. Castells, J. Pelkonen, and J. Monkkonen (2010)
J. Biol. Chem. 285, 1967-1979
 |  Abstract »  |  Full Text »  |  PDF »

CA-074Me Protection against Anthrax Lethal Toxin.

Z. L. Newman, S. H. Leppla, and M. Moayeri (2009)
Infect. Immun. 77, 4327-4336
 |  Abstract »  |  Full Text »  |  PDF »

Moderate exercise attenuates caspase-3 activity, oxidative stress, and inhibits progression of diabetic renal disease in db/db mice.

S. Ghosh, M. Khazaei, F. Moien-Afshari, L. S. Ang, D. J. Granville, C. B. Verchere, S. R. Dunn, P. McCue, A. Mizisin, K. Sharma, et al. (2009)
Am J Physiol Renal Physiol 296, F700-F708
 |  Abstract »  |  Full Text »  |  PDF »

Mitochondrial abnormalities in spinal and bulbar muscular atrophy.

S. Ranganathan, G. G. Harmison, K. Meyertholen, M. Pennuto, B. G. Burnett, and K. H. Fischbeck (2009)
Hum. Mol. Genet. 18, 27-42
 |  Abstract »  |  Full Text »  |  PDF »

Sensitization to the Lysosomal Cell Death Pathway by Oncogene-Induced Down-regulation of Lysosome-Associated Membrane Proteins 1 and 2.

N. Fehrenbacher, L. Bastholm, T. Kirkegaard-Sorensen, B. Rafn, T. Bottzauw, C. Nielsen, E. Weber, S. Shirasawa, T. Kallunki, and M. Jaattela (2008)
Cancer Res. 68, 6623-6633
 |  Abstract »  |  Full Text »  |  PDF »

Death Receptors and Caspases But Not Mitochondria Are Activated in the GDNF- or BDNF-Deprived Dopaminergic Neurons.

L.-y. Yu, M. Saarma, and U. Arumae (2008)
J. Neurosci. 28, 7467-7475
 |  Abstract »  |  Full Text »  |  PDF »

Phosphorylation of Murine Caspase-9 by the Protein Kinase Casein Kinase 2 Regulates Its Cleavage by Caspase-8.

M. A. McDonnell, Md. J. Abedin, M. Melendez, T. N. Platikanova, J. R. Ecklund, K. Ahmed, and A. Kelekar (2008)
J. Biol. Chem. 283, 20149-20158
 |  Abstract »  |  Full Text »  |  PDF »

Intracellular Fate Investigation of Bio-Eliminable Polymeric Nanoparticles by Confocal Laser Scanning Microscopy.

F. Chiellini, D. Dinucci, C. Bartoli, A. M. Piras, and E. Chiellini (2007)
Journal of Bioactive and Compatible Polymers 22, 667-685
 |  Abstract »  |  PDF »

Commitment to Apoptosis in CD4+ T Lymphocytes Productively Infected with Human Immunodeficiency Virus Type 1 Is Initiated by Lysosomal Membrane Permeabilization, Itself Induced by the Isolated Expression of the Viral Protein Nef.

M. Laforge, F. Petit, J. Estaquier, and A. Senik (2007)
J. Virol. 81, 11426-11440
 |  Abstract »  |  Full Text »  |  PDF »

Breast cancer cells with acquired antiestrogen resistance are sensitized to cisplatin-induced cell death.

C. W. Yde, M. Gyrd-Hansen, A. E. Lykkesfeldt, O.-G. Issinger, and J. Stenvang (2007)
Mol. Cancer Ther. 6, 1869-1876
 |  Abstract »  |  Full Text »  |  PDF »