Differential protection by wildtype vs. organelle-specific Bcl-2 suggests a combined requirement of both the ER and mitochondria in ceramide-mediated caspase-independent programmed cell death
https://doi.org/10.1186/1748-717X-4-41
© Deerberg et al; licensee BioMed Central Ltd. 2009
Received: 22 August 2009
Accepted: 9 October 2009
Published: 9 October 2009
Abstract
Background
Programmed cell death (PCD) is essential for development and homeostasis of multicellular organisms and can occur by caspase-dependent apoptosis or alternatively, by caspase-independent PCD (ciPCD). Bcl-2, a central regulator of apoptosis, localizes to both mitochondria and the endoplasmic reticulum (ER). Whereas a function of mitochondrial and ER-specific Bcl-2 in apoptosis has been established in multiple studies, corresponding data for ciPCD do not exist.
Methods
We utilized Bcl-2 constructs specifically localizing to mitochondria (Bcl-2 ActA), the ER (Bcl-2 cb5), both (Bcl-2 WT) or the cytosol/nucleus (Bcl-2 ΔTM) and determined their protective effect on ceramide-mediated ciPCD in transiently and stably transfected Jurkat cells. Expression of the constructs was verified by immunoblots. Ceramide-mediated ciPCD was induced by treatment with human recombinant tumor necrosis factor and determined by flow cytometric measurement of propidium iodide uptake as well as by optical analysis of cell morphology.
Results
Only wildtype Bcl-2 had the ability to efficiently protect from ceramide-mediated ciPCD, whereas expression of Bcl-2 solely at mitochondria, the ER, or the cytosol/nucleus did not prevent ceramide-mediated ciPCD.
Conclusion
Our data suggest a combined requirement for both mitochondria and the ER in the induction and the signaling pathways of ciPCD mediated by ceramide.
Keywords
Background
The survival and homeostasis of multicellular organisms critically depends on programmed cell death (PCD) to correctly remove cells that are potentially harmful or which have fulfilled their function. Although caspase-dependent apoptosis, the most well-studied form of PCD, represents the principal suicide program in most physiological settings [1], many cells have the ability to commit suicide by caspase-independent modes of PCD (ciPCD) [2]. ciPCD fulfills vital functions in processes such as the negative selection of lymphocytes, the termination of immune responses, embryonic removal of interdigital webs, regulation of bone growth, ovulation, and cellular turnover in the intestine [3]. Furthermore, ciPCD has been implicated the pathology of hyperacute shock [4], pancreatitis [5, 6], cerebral and myocardial ischemia-reperfusion injury, epilepsy, Alzheimer's disease and other inflammatory injuries, as well as in the destruction of cells by pathogens such as HIV, vaccinia virus, Shigella and Salmonella [3, 7]. In contrast to apoptosis, the molecular mechanisms of ciPCD are just beginning to be unraveled, e. g. by the recent identification of RIP3, enzymes of the energy metabolism, Nox1, CYLD, Bmf, and cathepsin and calpain proteases as candidate mediators of ciPCD [2, 8–10]. However, and despite these advances, a coherent picture of the molecular steps in ciPCD is still lacking.
Mitochondria have been identified not only as essential elicitors of apoptosis, but also of ciPCD, e.g. by release of proteins such as AIF, EndoG and HtrA2/OMI, as well as by production of reactive oxygen species (ROS) [2, 11–13]. It has been proposed that excess formation of ROS triggers ciPCD by activation of the DNA repair enzyme PARP, followed by intracellular depletion of NAD+ and ATP, nuclear translocation of AIF and finally, death [14]. Similar to mitochondria, the endoplasmic reticulum (ER) - as the main site for critical cellular functions such as protein folding, lipid biosynthesis, and calcium storage in the cell - has also been implicated in the induction of both apoptosis [15] and ciPCD [2, 12, 13]. In fact, the ER may play a key role in certain types of ciPCD, as intracellular calcium influx caused by ER stress induces activation of calpains, a family of calcium-dependent cytosolic proteases that can elicit ciPCD [2].
Members of the Bcl-2 family of proteins have long been recognized as central regulators of mitochondrial apoptosis, primarily by controlling the permeabilization of the outer mitochondrial membrane [15]. They have also been found localized at the ER, where they regulate apoptosis in response to a range of cellular stresses, and also in the nucleus [15], where they may fulfill yet unknown functions. Similar to the established protective functions of mitochondrial Bcl-2, expression of Bcl-2 at the ER has been shown to protect cells from apoptosis, e. g. by regulating the release of proapoptotic calcium, reducing calcium-uptake by mitochondria and subsequent calpain-dependent apoptosis [15]. The role of Bcl-2 in mitochondrial vs. ER-mediated apoptosis has been further addressed in a number of studies by the use of Bcl-2 constructs that specifically localize to mitochondria or to the ER [16–23]. In these studies, Bcl-2 localizing to the ER was shown to interfere with apoptosis induction by some (c-myc overexpression, etoposide, staurosporine, tunicamycin, brefeldin A, ceramide, ionizing radiation, thapsigargin, Bax, Bad) but not all stimuli (reviewed in [24, 25]).
In contrast to this relative wealth of available data on apoptosis, the role of ER- vs. mitochondrially localized Bcl-2 in ciPCD has not been investigated so far. Here, we utilize ciPCD elicited by ceramide as a model system, a lipid second messenger that has been recognized as important in radiation-induced elimination of tumor cells [26, 27]. We demonstrate for the first time that in contrast to wildtype Bcl-2, restricted expression of Bcl-2 solely at mitochondria-, the ER-, or the cytosol/nucleus is insufficient to prevent ciPCD. Therefore, our data suggest a combined requirement of mitochondria and the ER in ceramide-mediated ciPCD and implicate the existence of a molecular crosstalk between both organelles.
Methods
Reagents and constructs
Highly purified human recombinant TNF (hTNF) was supplied by BASF Bioresearch. Benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (zVAD-fmk) was obtained from Bachem. Cycloheximide (CHX) was purchased from Sigma. Expression constructs cloned in the vector pRc/CMV (Invitrogen) encoding wildtype Bcl-2 (Bcl-2 WT), cytosolic Bcl-2 lacking the transmembrane domain (Bcl-2 ΔTM) and Bcl-2 mutants restricted to the outer mitochondrial membrane (Bcl-2 ActA) or to the endoplasmic reticulum (Bcl-2 cb5) were kindly provided by B. Leber (McMaster University, Hamilton, Canada).
Cell culture and transfections
Wildtype human leukemic Jurkat cells were originally obtained from the American Type Culture Collection. Jurkat cells stably transfected with the plasmid pSFFV-Bcl-2, overexpressing full length human Bcl-2 at levels 10-20-fold over untransfected cells [28] were kindly provided by S. Korsmeyer (Harvard Medical School, Boston, USA). Cells were maintained in a mixture of Click's/RPMI 1640 (50/50% v/v) supplemented with 10% v/v FCS, 2 mM glutamine and 50 μg/ml each of streptomycin and penicillin in a humidified incubator containing 5% w/v CO2. Transient transfections of wildtype Jurkat cells were performed by Amaxa nucleofection (Lonza Cologne), using solution V and program C-16. Jurkat cells stably expressing the various versions of Bcl-2 in pRc/CMV were obtained by electroporation using a Gene pulser II (Bio-Rad) followed by selection with Geneticin (Invitrogen) and were used as pool transfectants.
Caspase assays
Cells were lysed in a buffer containing 10 mM Hepes pH 7.4, 142 mM KCl, 5 mM MgCl2, 1 mM EGTA, 0.2% v/v NP40, 1 mM DTT and 2 mM Pefabloc. To generate positive controls for activation, cytosolic extracts of untreated cells were equilibrated for 1 h at 30°C after the addition of 1 mM dATP and 10 μM cytochrome c to permit activation of caspases and subsequent cleavage of substrate proteins. To measure caspase activity, 100 μl of caspase buffer (20 mM Pipes, 100 mM NaCl, 10 mM DTT, 1 mM EDTA, 0.1% w/v CHAPS, 10% w/v sucrose, pH 7.2) containing 100 μM zDEVD-afc or zIETD-afc (Calbiochem) were added to 5 μl of cytosolic extract (50 μg protein) and incubated at 37°C. The release of afc was measured as emission at 505 nm upon excitation at 405 nm using a Labsystems Fluoroskan II fluorimeter equipped with a thermostated plate reader.
Microscopy
For documentation of cell morphology, images from unfixed cells were obtained using an Axiovert 100 microscope (Zeiss) and a DS-5 M camera (Nikon).
Cytotoxicity assays
For flow cytometric measurement of cell death, cells were seeded in six-well plates at 5 × 105 cells/well. Following treatment, cells were collected by centrifugation and resuspended in PBS/5 mM EDTA containing 2 μg/ml propidium iodide (PI), and the red fluorescence was measured on a FACSCalibur flow cytometer (BD Biosciences). Since we observed that measurement exclusively of PI-positive cells did not account for a significant fraction of highly disintegrated dead cells that - due to diffusion of the dye - had already become PI-negative again (shown e. g. in Fig. 1C, right panels, lower left quadrants), we alternatively measured the fraction of large, PI negative cells (lower right quadrants) which represents viable, intact cells. For each measurement, a total of 10.000 cells was analyzed.
Immunoblots
Cells were collected and lysed in TNE buffer (50 mM Tris pH 8.0, 150 mM NaCl, 1% v/v NP40, 2 mM EDTA) containing 10 μg/ml pepstatin/aprotinin/leupeptin, 1 mM sodium orthovanadate and 5 mM NaF. After removal of insoluble material by centrifugation at 10.000 × g and 4°C for 15 min, the protein concentration of the supernatants was measured using a BCA assay (Pierce). 30 μg of cell protein per lane were resolved by electrophoresis on 12.5% w/v SDS polyacrylamide gels (SDS-PAGE). After electrophoretic transfer to Protran nitrocellulose (Whatman), reactive proteins were detected using a monoclonal IgG1 antibody specific for human Bcl-2 (sc-7382, Santa Cruz) and the ECL detection kit (GE Healthcare).
Results
Wildtype Bcl-2 protects from ceramide-mediated ciPCD
Wildtype Bcl-2 protects from ceramide-induced ciPCD. (A) Activity of caspase-8 and -3 in wildtype Jurkat cells in response to TNF/CHX/zVAD inducing ciPCD or TNF/CHX as a proapoptotic stimulus. Cells were incubated with 100 ng/ml hTNF in combination with 2 μg/ml CHX and/or 50 μM zVAD-fmk for 4 h before activation of caspases -8 and -3 was determined by measuring the cleavage of fluorogenic substrates (zIETD-afc and zDEVD-afc) over 120 minutes. Prior to stimulation, cells were preincubated with 50 μM zVAD-fmk for 30 min (for stimulations with TNF/CHX/zVAD) or medium (stimulations with TNF/CHX). For positive control, caspases were activated in vitro by adding cytochrome c and dATP (Cyt c/dATP) to the cell extracts. (B) Wildtype (untransfected) Jurkat cells and Jurkat cells overexpressing pSFFV-Bcl-2 (Bcl-2 WT) were left untreated or stimulated with 100 ng/ml hTNF in combination with 5 μg/ml CHX and 50 μM zVAD-fmk for 20 h before micrographs of the cells were taken to document their morphology. Prior to stimulation, the cells were preincubated for 60 min with 50 μM zVAD-fmk. As representative examples, one healthy cell and one cell undergoing ciPCD with necrosis-like morphology are marked by a black or a white arrow, respectively. (C) In parallel, uptake of PI was determined by flow cytometry as a marker for loss of plasma membrane integrity (see „Materials and Methods‟). The percentage of viable cells (PI-negative, large) is indicated in the lower right quadrants of the dot plots. One representative experiment out of three performed is shown. (D) Quantification of cell viability data. The bar graphs represent the means from all three independent experiments, error bars indicate the respective standard deviations.
Transient expression of Bcl-2 constructs with restricted subcellular localization
Transient expression of Bcl-2 constructs with defined subcellular localization in Jurkat cells. (A) Schematic representation of human wildtype Bcl-2 (Bcl-2 WT) localizing to both mitochondria and the ER, Bcl-2 ActA targeted to mitochondria, Bcl-2 cb5 expressed at the ER, and Bcl-2 ΔTM localized in the cytosol. The Bcl-2 homology domains BH1-BH4 are indicated together with the transmembrane domain (TM) for wildtype Bcl-2, which is replaced by amino acids from Listeria monocytogenes ActA, from rat cytochrome b5 (cb5) or deleted altogether in the other constructs. (B) amino acid sequence of the carboxyterminus of wildtype Bcl-2, Bcl-2 ActA, Bcl-2 cb5 and Bcl-2 ΔTM. The amino acids derived from ActA and cb5 are shown in bold. (C) Wildtype Jurkat cells were transiently nucleofected with empty vector pRc/CMV or with pRc/CMV encoding wildtype Bcl-2. 24 h after transfection, the cells were stimulated with 100 ng/ml hTNF in combination with 5 μg/ml CHX and 50 μM zVAD-fmk or left untreated for another 24 h. Prior to stimulation, the cells were preincubated for 60 min with 50 μM zVAD-fmk. PI-uptake was determined by flow cytometry and the percentage of viable cells is indicated in the lower right quadrants of the dot plots. One representative experiment out of three performed is shown. (D) Quantification of cell viability data. The bar graphs represent the means from all three independent experiments, error bars indicate the respective standard deviations. Due to the transfection procedure, the cells generally display a lower viability than untransfected cells (Fig. 1). (E) In parallel, expression of Bcl-2 in the transfectants was visualized by Western blot analysis. The band in vector transfectants represents endogenous Bcl-2.
Wildtype, but not organelle-specific Bcl-2 protects from ceramide-mediated ciPCD in stably transfected Jurkat cells
Wildtype, but not organelle-restricted Bcl-2 protects stably transfected Jurkat cells from ceramide-mediated ciPCD. (A) Expression of pRc/CMV-encoded Bcl-2 WT, Bcl-2 cb5, Bcl-2 ActA and Bcl-2 ΔTM-constructs in stably transfected Jurkat cells. Cell lysates were prepared from untransfected Jurkat cells or Jurkat cells stably transfected with empty vector or with Bcl-2 constructs targeted to the ER, mitochondria, both, or the cytosol as indicated. Expression of the constructs was verified by immunoblot with a Bcl-2-specific antibody (sc-7382, Santa Cruz). Multiple bands result from detection of the endogenous Bcl-2 protein in addition to the construct (see untransfected Jurkat cells). (B) Flow cytometric analysis of PI-uptake in untransfected and stably transfected Jurkat cells. Prior to stimulation, the cells were preincubated for 60 min with 50 μM zVAD-fmk. After that, ceramide-mediated ciPCD was induced by treatment with 100 ng/ml hTNF in combination with 5 μg/ml CHX and 50 μM zVAD-fmk for 48 h, or the cells were left completely untreated. The percentage of viable cells is shown in the lower right quadrants of the dot plots. (C) Bar graphs showing the fraction of viable cells for each of the stably transfected lines depicted in (B). Similar results were obtained in two additional experiments with different incubation times (24 and 72 h), although with increased or reduced overall viability (data not shown).
Discussion
In this study, we show for the first time that Bcl-2-mediated protection from non-apoptotic, caspase-independent PCD requires the full-length, wildtype protein whereas a limited expression of Bcl-2 at mitochondria, the ER, or the cytosol/nucleus is not sufficient to prevent ciPCD elicited by ceramide. The failure of the organelle-specific constructs to confer protection is not due to an inefficient level of expression (Fig. 3A) or an incorrect localization [21], and their functionality has been previously demonstrated for apoptotic PCD using the same stable transfectants as analyzed here [21, 22]. Moreover, the protection by wildtype Bcl-2 was independently seen in two distinct stably transfected Jurkat lines, expressed from different vector backbones (pSFFV-neo, pRc/CMV).
The fact that expression of Bcl-2 restricted to mitochondria, the ER (or the cytosol/nucleus, if the results with Bcl-2 ΔTM are additionally taken into account) is not effectively suppressing ceramide-induced ciPCD may suggest that wildtype Bcl-2 interferes with this form of PCD at yet another cellular site. However, this is unlikely, as wildtype Bcl-2 has been detected only at mitochondria, the ER and the nucleus in numerous previous studies (reviewed in [15, 35]). Alternatively, the exclusive protection by wildtype Bcl-2 may be explained by a combined requirement of Bcl-2 at both mitochondria and at the ER, suggesting that both organelles coordinately participate in ciPCD via a molecular crosstalk. This hypothesis appears especially attractive because a similar crosstalk has been shown for apoptotic PCD, where the ER controls mitochondrial apoptosis by releasing proapoptotic calcium [15]. Although calcium likewise appears as an attractive candidate molecule for ER-mitochondrial crosstalk in ciPCD (especially when considering that the calcium-dependent calpain proteases can elicit ciPCD [2]), the precise nature of this crosstalk will have to be clarified in future studies.
Notably, in autophagy, a distinct form of ciPCD, Bcl-2 has been recognized to act as a negative regulator specifically at the ER by binding to the autophagy-inducing protein Beclin-1 [15].
Even though the role of Bcl-2 has been most intensively studied in mitochondria and the ER, Bcl-2 also localizes to the nucleus. However, apart from a recent study where it was shown that nuclear Bcl-2 forms an integral part of mitotic chromosomes [36], information on the functions of nuclear Bcl-2 are scarce. Therefore, nuclear functions of Bcl-2 may likewise be required for suppression of ciPCD in concert with its mitochondrial and ER-specific functions.
In summary, the further exploration of the exact roles of mitochondrial, ER- and nucleus-specific Bcl-2 as well as their potential crosstalk will provide further insight into the molecular mechanisms by which ceramide-mediated ciPCD is executed. At the clinical level, this may directly prove beneficial also for radiation therapy by providing novel options to eliminate tumor cells that have become resistant to apoptotic death signals.
Conclusion
Overview scheme depicting the proposed requirement of the ER and mitochondria in TNF-R1/ceramide-mediated ciPCD. Known proximal mediators of TNF-R1/ceramide-induced ciPCD are indicated, as are mediators of ciPCD that potentially act downstream of the ER and mitochondria. Only wildtype Bcl-2 simultaneously acting at the ER, at mitochondria and at the nucleus efficiently blocks the caspase-independent death signals of TNF-R1/ceramide, whereas Bcl-2 constructs specifically localizing to each organelle do not prevent ciPCD. This suggests that the corresponding signaling pathways of TNF-R1/ceramide target both the ER and mitochondria, and that both organelles participate in ciPCD via a molecular crosstalk. The nucleus may represent a further organelle that participates in these signaling pathways, yet its role remains to be confirmed.
Declarations
Acknowledgements
We thank D. Kabelitz for his continuous support and encouragement. This work was supported by grants from the from the Forschungskommission der Medizinischen Fakultät and from the Deutsche Forschungsgemeinschaft to D. A. (SFB 415, A4). The authors have no conflicting financial interests.
Authors’ Affiliations
References
- Degterev A, Yuan J: Expansion and evolution of cell death programmes. Nat Rev Mol Cell Biol 2008, 9: 378-390. 10.1038/nrm2393View ArticlePubMedGoogle Scholar
- Bröker LE, Kruyt FA, Giaccone G: Cell death independent of caspases: a review. Clin Cancer Res 2005, 11: 3155-3162. 10.1158/1078-0432.CCR-04-2223View ArticlePubMedGoogle Scholar
- Festjens N, Vanden Berghe T, Vandenabeele P: Necrosis, a well-orchestrated form of cell demise: signalling cascades, important mediators and concomitant immune response. Biochim Biophys Acta 2006, 1757: 1371-1387. 10.1016/j.bbabio.2006.06.014View ArticlePubMedGoogle Scholar
- Cauwels A, Janssen B, Waeytens A, Cuvelier C, Brouckaert P: Caspase inhibition causes hyperacute tumor necrosis factor-induced shock via oxidative stress and phospholipase A2. Nat Immunol 2003, 4: 387-393. 10.1038/ni914View ArticlePubMedGoogle Scholar
- He S, Wang L, Miao L, Wang T, Du F, Zhao L, Wang X: Receptor interacting protein kinase-3 determines cellular necrotic response to TNF-alpha. Cell 2009, 137: 1100-1111. 10.1016/j.cell.2009.05.021View ArticlePubMedGoogle Scholar
- Zhang DW, Shao J, Lin J, Zhang N, Lu BJ, Lin SC, Dong MQ, Han J: RIP3, an energy metabolism regulator that switches TNF-induced cell death from apoptosis to necrosis. Science 2009, 325: 332-336. 10.1126/science.1172308View ArticlePubMedGoogle Scholar
- Cho YS, Challa S, Moquin D, Genga R, Ray TD, Guildford M, Chan FK: Phosphorylation-driven assembly of the RIP1-RIP3 complex regulates programmed necrosis and virus-induced inflammation. Cell 2009, 137: 1112-1123. 10.1016/j.cell.2009.05.037PubMed CentralView ArticlePubMedGoogle Scholar
- Declercq W, Vanden Berghe T, Vandenabeele P: RIP kinases at the crossroads of cell death and survival. Cell 2009, 138: 229-232. 10.1016/j.cell.2009.07.006View ArticlePubMedGoogle Scholar
- Kim YS, Morgan MJ, Choksi S, Liu ZG: TNF-induced activation of the Nox1 NADPH oxidase and its role in the induction of necrotic cell death. Mol Cell 2007, 26: 675-687. 10.1016/j.molcel.2007.04.021View ArticlePubMedGoogle Scholar
- Yazdanpanah B, Wiegmann K, Tchikov V, Krut O, Pongratz C, Schramm M, Kleinridders A, Wunderlich T, Kashkar H, Utermöhlen O, Brüning JC, Schütze S, Krönke M: Riboflavin kinase couples TNF receptor 1 to NADPH oxidase. Nature 2009,460(7259):1159-1163. 10.1038/nature08206View ArticlePubMedGoogle Scholar
- Chipuk JE, Green DR: Do inducers of apoptosis trigger caspase-independent cell death? Nat Rev Mol Cell Biol 2005, 6: 268-275. 10.1038/nrm1573View ArticlePubMedGoogle Scholar
- Leist M, Jäättelä M: Four deaths and a funeral: from caspases to alternative mechanisms. Nat Rev Mol Cell Biol 2001, 2: 589-598. 10.1038/35085008View ArticlePubMedGoogle Scholar
- Jäättelä M, Tschopp J: Caspase-independent cell death in T lymphocytes. Nat Immunol 2003, 4: 416-423. 10.1038/ni0503-416View ArticlePubMedGoogle Scholar
- Hong SJ, Dawson TM, Dawson VL: Nuclear and mitochondrial conversations in cell death: PARP-1 and AIF signaling. Trends Pharmacol Sci 2004, 25: 259-264. 10.1016/j.tips.2004.03.005View ArticlePubMedGoogle Scholar
- Szegezdi E, Macdonald DC, Ni Chonghaile T, Gupta S, Samali A: Bcl-2 family on guard at the ER. Am J Physiol Cell Physiol 2009, 296: C941-953. 10.1152/ajpcell.00612.2008View ArticlePubMedGoogle Scholar
- Zhu W, Cowie A, Wasfy GW, Penn LZ, Leber B, Andrews DW: Bcl-2 mutants with restricted subcellular location reveal spatially distinct pathways for apoptosis in different cell types. EMBO J 1996, 15: 4130-4141.PubMed CentralPubMedGoogle Scholar
- Froesch BA, Aimé-Sempé C, Leber B, Andrews D, Reed JC: Inhibition of p53 transcriptional activity by Bcl-2 requires its membrane-anchoring domain. J Biol Chem 1999, 274: 6469-6475. 10.1074/jbc.274.10.6469View ArticlePubMedGoogle Scholar
- Häcki J, Egger L, Monney L, Conus S, Rossé T, Fellay I, Borner C: Apoptotic crosstalk between the endoplasmic reticulum and mitochondria controlled by Bcl-2. Oncogene 2000, 19: 2286-2295. 10.1038/sj.onc.1203592View ArticlePubMedGoogle Scholar
- Annis MG, Zamzami N, Zhu W, Penn LZ, Kroemer G, Leber B, Andrews DW: Endoplasmic reticulum localized Bcl-2 prevents apoptosis when redistribution of cytochrome c is a late event. Oncogene 2001, 20: 1939-1952. 10.1038/sj.onc.1204288View ArticlePubMedGoogle Scholar
- Soucie EL, Annis MG, Sedivy J, Filmus J, Leber B, Andrews DW, Penn LZ: Myc potentiates apoptosis by stimulating Bax activity at the mitochondria. Mol Cell Biol 2001, 21: 4725-4736. 10.1128/MCB.21.14.4725-4736.2001PubMed CentralView ArticlePubMedGoogle Scholar
- Rudner J, Lepple-Wienhues A, Budach W, Berschauer J, Friedrich B, Wesselborg S, Schulze-Osthoff K, Belka C: Wild-type, mitochondrial and ER-restricted Bcl-2 inhibit DNA damage-induced apoptosis but do not affect death receptor-induced apoptosis. J Cell Sci 2001, 114: 4161-4172.PubMedGoogle Scholar
- Handrick R, Rudner J, Müller I, Eibl H, Belka C, Jendrossek V: Bcl-2 mediated inhibition of erucylphosphocholine-induced apoptosis depends on its subcellular localisation. Biochem Pharmacol 2005, 70: 837-850. 10.1016/j.bcp.2005.06.021View ArticlePubMedGoogle Scholar
- Bhatt K, Feng L, Pabla N, Liu K, Smith S, Dong Z: Effects of targeted Bcl-2 expression in mitochondria or endoplasmic reticulum on renal tubular cell apoptosis. Am J Physiol Renal Physiol 2008, 294: F499-507. 10.1152/ajprenal.00415.2007View ArticlePubMedGoogle Scholar
- Rudner J, Jendrossek V, Belka C: New insights in the role of Bcl-2 Bcl-2 and the endoplasmic reticulum. Apoptosis 2002, 7: 441-447. 10.1023/A:1020087108926View ArticlePubMedGoogle Scholar
- Thomenius MJ, Distelhorst CW: Bcl-2 on the endoplasmic reticulum: protecting the mitochondria from a distance. J Cell Sci 2003, 116: 4493-4499. 10.1242/jcs.00829View ArticlePubMedGoogle Scholar
- Weichselbaum R: Radiation's outer limits. Nat Med 2005, 11: 477-478. 10.1038/nm0505-477View ArticlePubMedGoogle Scholar
- Kolesnick R, Fuks Z: Radiation and ceramide-induced apoptosis. Oncogene 2003, 22: 5897-5906. 10.1038/sj.onc.1206702View ArticlePubMedGoogle Scholar
- Armstrong RC, Aja T, Xiang J, Gaur S, Krebs JF, Hoang K, Bai X, Korsmeyer SJ, Karanewsky DS, Fritz LC, Tomaselli KJ: Fas-induced activation of the cell death-related protease CPP32 is inhibited by Bcl-2 and by ICE family protease inhibitors. J Biol Chem 1996, 271: 16850-16855. 10.1074/jbc.271.9.4978View ArticlePubMedGoogle Scholar
- Strelow A, Bernardo K, Adam-Klages S, Linke T, Sandhoff K, Krönke M, Adam D: Overexpression of acid ceramidase protects from tumor necrosis factor-induced cell death. J Exp Med 2000, 192: 601-611. 10.1084/jem.192.5.601PubMed CentralView ArticlePubMedGoogle Scholar
- Lange A, Thon L, Mathieu S, Adam D: The apoptosis inhibitory domain of FE65-like protein 1 regulates both apoptotic and caspase-independent programmed cell death mediated by tumor necrosis factor. Biochem Biophys Res Commun 2005, 335: 575-583. 10.1016/j.bbrc.2005.07.125View ArticlePubMedGoogle Scholar
- Thon L, Möhlig H, Mathieu S, Lange A, Bulanova E, Winoto-Morbach S, Schütze S, Bulfone-Paus S, Adam D: Ceramide mediates caspase-independent programmed cell death. FASEB J 2005, 19: 1945-1956. 10.1096/fj.05-3726comView ArticlePubMedGoogle Scholar
- Thon L, Mathieu S, Kabelitz D, Adam D: The murine TRAIL receptor signals caspase-independent cell death through ceramide. Exp Cell Res 2006, 312: 3808-3821. 10.1016/j.yexcr.2006.08.017View ArticlePubMedGoogle Scholar
- Hennet T, Bertoni G, Richter C, Peterhans E: Expression of Bcl-2 protein enhances the survival of mouse fibrosarcoid cells in tumor necrosis factor-mediated cytotoxicity. Cancer Res 1993, 53: 1456-1460.PubMedGoogle Scholar
- Denecker G, Vercammen D, Steemans M, Berghe T, Brouckaert G, van Loo G, Zhivotovsky B, Fiers W, Grooten J, Declercq W, Vandenabeele P: Death receptor-induced apoptotic and necrotic cell death: differential role of caspases and mitochondria. Cell Death Differ 2001, 8: 829-840. 10.1038/sj.cdd.4400883View ArticlePubMedGoogle Scholar
- Annis MG, Yethon JA, Leber B, Andrews DW: There is more to life and death than mitochondria: Bcl-2 proteins at the endoplasmic reticulum. Biochim Biophys Acta 2004, 1644: 115-123. 10.1016/j.bbamcr.2003.07.001View ArticlePubMedGoogle Scholar
- Barboule N, Demeter K, Benmeradi N, Larminat F: Bcl-2 is an integral component of mitotic chromosomes. Cell Biol Int 2009, 33: 572-577. 10.1016/j.cellbi.2009.02.012View ArticlePubMedGoogle Scholar
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