Induction of a Common Pathway of Apoptosis by Staurosporine

Bertrand, Richard; Solary, Éric; O’Connor, Patrick M.; Kohn, Kurt W.; Pommier, ﻿Yves

doi:10.1006/excr.1994.1093

Cited by 466 publications

(342 citation statements)

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“…Fas is the cell-surface receptor for Fas ligand and is implicated in various physiological and pathological processes (Nagata and Goldstein, 1995;Abbas, 1996;Boise and Thompson, 1996;Nagata, 1996). Staurosporine is a broad-spectrum protein kinase inhibitor that is able to induce apoptosis in various types of cells (Bertrand et al, 1994;Jacobson et al, 1996). Jurkat T cells treated with anti-Fas antibody (Figure 1a) or staurosporine ( Figure 1b) underwent apoptotic cell death within a few hours as described previously .…”

Section: Stepwise Activation Of Caspases In Apoptotic Jurkat Cells Rementioning

confidence: 62%

Affinity labeling displays the stepwise activation of ICE-related proteases by Fas, staurosporine, and CrmA-sensitive caspase-8

et al. 1997

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The activation of multiple interleukin-1b converting enzyme-related proteases (caspases) in apoptotic mammalian cells raises questions as to whether the multiple active caspases have distinct roles in apoptotic execution as well as how these proteases are organized in apoptotic signaling pathways. Here we used an a nity-labeling agent, YV(bio)KD-aomk, to investigate the caspases activated during apoptotic cell death. YV(bio)KD-aomk identi®ed six distinct polypeptides corresponding to active caspases in Fas-stimulated Jurkat T cells. On staurosporine treatment, four polypeptides were detected. Competition experiments showed that the labeled caspases have distinct substrate preferences. Stepwise appearance of the labeled caspases in each cell death event was consistent with the view that the activated caspases are organized into protease cascades. Moreover, we found that stepwise activation of caspases similar to that induced by Fas ligation is triggered by exposing non-apoptotic Jurkat cell extracts to caspase-8 (MACH/FLICE/Mch5). Conversely, CrmA protein, a viral suppressor of Fas-induced apoptosis, inhibited the protease activity of caspase-8. Overall, these ®ndings provide evidence that caspase-8, a CrmA-sensitive protease, is responsible for initiating the stepwise activation of multiple caspases in Fas-stimulated cells.

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Section: Stepwise Activation Of Caspases In Apoptotic Jurkat Cells Rementioning

confidence: 62%

Affinity labeling displays the stepwise activation of ICE-related proteases by Fas, staurosporine, and CrmA-sensitive caspase-8

et al. 1997

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“…Using STS, we have examined if cells lacking cyt c experience normal apoptotic signalling upstream of the mitochondria, and whether the release of other apoptogenic IMS proteins such as Smac/DIABLO occurs in the absence of cyt c. We report here that proapoptotic signalling at the mitochondria still occurs in the cyt cÀ/À cells, as Bax is recruited to the mitochondria in the presence of STS. However, we found that Smac/DIABLO is not released from the mitochondria of cyt cÀ/À cells in response to STS (a protein kinase inhibitor 7 ) or etoposide (a topoisomerase inhibitor 8 ). This finding indicates that there are pleiotropic defects in apoptotic signalling effected by mitochondria in cyt cÀ/À cells and raises the interesting possibility that release of Smac/DIABLO from mitochondria is dependent on either the presence of cyt c, as such, or its prior mobilisation from mitochondria to cytosol.…”

Section: Introductionmentioning

confidence: 72%

Smac/DIABLO is not released from mitochondria during apoptotic signalling in cells deficient in cytochrome c

2005

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We have characterised the apoptotic defects in cells null for cytochrome c (cyt cÀ/À). Such cells treated with staurosporine (STS) exhibited translocation to the mitochondria and activation of the proapoptotic signalling molecule Bax but failed to release Smac/DIABLO from these organelles, judged by both confocal microscopy and Western blotting. While reference cells expressing cytochrome c released both it and Smac/DIABLO under a variety of conditions of apoptotic induction, we have never observed release of Smac/DIABLO from cyt cÀ/À cells. We eliminate the possibility that proteasomal degradation of cytosolically localised Smac/ DIABLO is responsible for our failure to visualise the protein outside the mitochondria. Our findings indicate an unanticipated nexus between release of cytochrome c and Smac/DIABLO from mitochondria, previously thought to be a more or less synchronised event early in apoptosis. We suggest that the failure of cyt cÀ/À cells to release Smac/DIABLO after recruitment of Bax to mitochondria represents an extreme manifestation of some inherent difference in the regulation of release of these two proteins from mitochondria.

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“…PMN were prepared and stained as above, but without adding propidium iodide. As a positive control, an extra group of PMN was incubated with staurosporine (200 nM, Sigma), a specific inducer of apoptosis [2]. After staining, cells were washed, and fixed with 2% paraformaldehyde for 1 h at room temperature.…”

Section: Phosphatidylserine Translocationmentioning

confidence: 99%

Tilmicosin-induced bovine neutrophil apoptosis is cell-specific and downregulates spontaneous LTB4 synthesis without increasing Fas expression

et al. 2004

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-The pathology of bacterial pneumonia, such as seen in the bovine lung infected with Mannheimia haemolytica, is due to pathogen virulence factors and to inflammation initiated by the host. Tilmicosin is a macrolide effective in treating bacterial pneumonia and recent findings suggest that this antibiotic may provide anti-inflammatory benefits by inducing polymorphonuclear neutrophilic leukocyte (PMN) apoptosis. Using an in vitro bovine system, we examined the cellspecificity of tilmicosin, characterized the changes in spontaneous leukotriene B 4 (LTB 4 ) synthesis by PMN exposed to the macrolide, and assessed its effects on PMN Fas expression. Previous findings demonstrated that tilmicosin is able to induce PMN apoptosis. These results were confirmed in this study by the Annexin-V staining of externalized phosphatidylserine and the analysis with flow cytometry. The cell-specificity of tilmicosin was assessed by quantification of apoptosis in bovine PMN, mononuclear leukocytes, monocyte-derived macrophages, endothelial cells, epithelial cells, and fibroblasts cultured with the macrolide. The effect of tilmicosin on spontaneous LTB 4 production by PMN was evaluated via an enzyme-linked immunosorbent assay. Finally, the mechanisms of tilmicosin-induced PMN apoptosis were examined by assessing the effects of tilmicosin on surface Fas expression on PMN. Tilmicosin-induced apoptosis was found to be at least partially cell-specific, as PMN were the only cell type tested to die via apoptosis in response to incubation with tilmicosin. PMN incubated with tilmicosin under conditions that induce apoptosis spontaneously produced less LTB 4 , but did not exhibit altered Fas expression. In conclusion, tilmicosin-induced apoptosis is specific to PMN, inhibits spontaneous LTB 4 production, and occurs through a pathway independent of Fas upregulation. macrolide / neutrophil / apoptosis / pasteurellosis / inflammation

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Induction of a Common Pathway of Apoptosis by Staurosporine

Cited by 466 publications

References 0 publications

Affinity labeling displays the stepwise activation of ICE-related proteases by Fas, staurosporine, and CrmA-sensitive caspase-8

Affinity labeling displays the stepwise activation of ICE-related proteases by Fas, staurosporine, and CrmA-sensitive caspase-8

Smac/DIABLO is not released from mitochondria during apoptotic signalling in cells deficient in cytochrome c

Tilmicosin-induced bovine neutrophil apoptosis is cell-specific and downregulates spontaneous LTB4 synthesis without increasing Fas expression

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