Ordering the Cytochrome c–initiated Caspase Cascade: Hierarchical Activation of Caspases-2, -3, -6, -7, -8, and -10 in a Caspase-9–dependent Manner

Slee, Elizabeth A.; Harte, Mary T.; Kluck, Ruth M.; Wolf, Beni B.; Casiano, Carlos A.; Newmeyer, Donald D.; Wang, Hong Gang; Reed, John C.; Nicholson, Donald W.; Alnemri, Emad S.; Green, Douglas R.; Martin, Séamus J.

doi:10.1083/jcb.144.2.281

Cited by 1,745 publications

(1,471 citation statements)

References 65 publications

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“…Our observations are also supported by recent in vitro studies, in which it was shown that caspase-9 cleaves directly caspase-3 and -7, but is unable to process caspase-6 (Srinivasula et al, 1998). In addition, depletion of caspase-3 from cell extracts abolished cytochrome c-induced processing of caspase-8 (Slee et al, 1999). In agreement with these in vitro data, we suggest that also in intact cells caspase-8 is activated downstream of caspase-9, most likely following caspase-3-mediated activation of caspase-6 (Figure 7).…”

Section: Discussionsupporting

confidence: 90%

“…Several lines of evidence support the idea that caspase-9 is the essential caspase in many forms of apoptosis (Li et al, 1997b;Kuida et al, 1998;Hakem et al, 1998;Sun et al, 1999). In addition, although caspase-8 has been initially suggested to interact with Apaf-1, a more detailed analysis showed that caspase-9 is the only Apaf-1 interacting protease in living cells (Slee et al, 1999). In accordance, our data demonstrate that overexpression of a dominant-negative caspase-9 mutant strongly inhibited both the cleavage of endogenous caspase-9, and -6 as well as of Bid and caspase-8 following drug treatment.…”

Section: Discussionmentioning

confidence: 87%

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Caspase-8/FLICE functions as an executioner caspase in anticancer drug-induced apoptosis

et al. 2000

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Caspase-8 plays an essential role in apoptosis triggered by death receptors. Through the cleavage of Bid, a proapoptotic Bcl-2 member, it further activates the mitochondrial cytochrome c/Apaf-1 pathway. Because caspase-8 can be processed also by anticancer drugs independently of death receptors, we investigated its exact role and order in the caspase cascade. We show that in Jurkat cells either de®cient for caspase-8 or overexpressing its inhibitor c-FLIP apoptosis mediated by CD95, but not by anticancer drugs was inhibited. In the absence of active caspase-8, anticancer drugs still induced the processing of caspase-9, -3 and Bid, indicating that Bid cleavage does not require caspase-8. Overexpression of Bcl-x L prevented the processing of caspase-8 as well as caspase-9, -6 and Bid in response to drugs, but was less e ective in CD95-induced apoptosis. Similar responses were observed by overexpression of a dominant-negative caspase-9 mutant. To further determine the order of caspase-8 activation, we employed MCF7 cells lacking caspase-3. In contrast to caspase-9 that was cleaved in these cells, anticancer drugs induced caspase-8 activation only in caspase-3 transfected MCF7 cells. Thus, our data indicate that, unlike its proximal role in receptor signaling, in the mitochondrial pathway caspase-8 rather functions as an amplifying executioner caspase.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 87%

Caspase-8/FLICE functions as an executioner caspase in anticancer drug-induced apoptosis

et al. 2000

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“…Bcl-2 and C9DN expression prevented the processing of procaspase 2 induced by anti-Fas and the agonist RRM MX2870-1, indicating that effector caspases were probably responsible for the activation of caspase 2. 13 Interestingly, the antagonist MX781 induced caspase 2 cleavage in Bcl-2-and C9DN-expressing cells to levels comparable to those seen in Jurkat-pcDNA cells (Figure 5a), suggesting an apical role for caspase 2 in MX781-induced apoptosis. 17 Slightly higher levels of procaspase 2 were observed in untreated Jurkat-Bcl-2 cells, which were not due to differences in protein loading (see Figure 4a and data not shown).…”

Section: Expression Of Bcl-2 and Bcl-x L In Jurkat Cells Prevents Rrmmentioning

confidence: 62%

“…17 This contrasts with the requirement of caspase 9 activity for the efficient cleavage of procaspase 2 by MX2870-1 and Fas signals. The latter resembles the activation of caspase 2 by effector caspases upon activation of the mitochondrial pathway, 13 as observed in UV-and TNF-dependent apoptosis. 66 Despite the activation of caspase 2 by MX781 in cells overexpressing Bcl-2 or C9DN, this is not sufficient to cause the rapid induction of DEVDase activity that is observed in control cells, in which the mitochondria-caspase 9 loop is functional.…”

Section: Discussionmentioning

confidence: 76%

“…[4][5][6][7][8] Released cytochrome c, along with Apaf-1 and procaspase 9, forms the apoptosome, 9,10 causing the proteolytic autoactivation of caspase 9, which in turn cleaves and activates effector caspases 3 and 7. [11][12][13][14] Smac release is necessary for the effective activation of caspases through interactions with inhibitor of apoptosis proteins (IAPs). Recent evidence suggests that caspase 2 activation upstream of the mitochondria can provoke the release of mitochondrial proapoptotic factors and is essential for UV-and stress-induced apoptosis.…”

Section: Introductionmentioning

confidence: 99%

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Retinoid-related molecules require caspase 9 for the effective release of Smac and the rapid induction of apoptosis

López-Hernández¹,

Ortiz²,

Bayón³

et al. 2003

Cell Death Differ

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Certain retinoid-related molecules (RRMs) with agonist or antagonist activities have been described to induce apoptosis in a variety of cancer cell lines and show promise for the treatment of cancer. Similar to other chemotherapeutic drugs, these retinoid analogs have been suggested to induce apoptosis through the intrinsic pathway, which requires the release of cytochrome c from the mitochondria for the effective activation of caspase 9. Expression of a catalytically inactive form of caspase 9, which functions as a dominant negative mutant, inhibits the induction of DEVDase activity and nuclear fragmentation by selective RRMs. Whereas the RRMs could induce the release of cytochrome c in the absence of caspase 9 activity, the later is necessary for the effective release of Smac/Diablo from the mitochondria. Furthermore, overexpression of Bcl-2 or Bcl-X L also inhibits RRM-induced apoptosis. We demonstrate that activation of caspase 2 by the agonist MX2870-1 requires caspase 9 activity and is inhibited by Bcl-2 overexpression. In contrast, the antagonist MX781 induces cleavage of procaspase 2 upstream of mitochondria and independently of caspase 9. Thus, two retinoid analogs with unique characteristics activate two distinct apical caspases (2 or 9) to initiate apoptosis. In addition to caspase-mediated cell death, sustained exposure to the RRMs can also lead to loss of cell viability in cells lacking caspase 9 activity or in cells stimulated in the presence of the caspase inhibitor Z-VADfmk. Moreover, MX2870-1 and MX781 produce cell cycle arrest independently of caspase activity and the retinoid receptors.

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Overexpression of Akt (protein kinase B) confers protection against apoptosis and prevents formation of ceramide in response to pro-apoptotic stimuli

et al. 1999

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An immortalized dorsal root ganglion cell line F-11 exhibits many properties of spinal cord neurons and undergoes apoptosis in response to growth factor withdrawal and the exogenous addition of inhibitors of phosphatidylinositol-3-kinase (PI3K). To elucidate the mechanism of apoptosis we generated F-11 clones which overexpressed either the p110 subunit of PI3K, a constitutively active form of protein kinase B/Akt (Myristoylated Akt), or a dominant-negative form (c-Akt). The first two constructs were protective against apoptosis induced by PI3K inhibitors such as wortmannin and LY294002. Caspase-3 (CPP32) levels peaked at 4 hr to 6 hr in response to pro-apoptotic drugs, and this increase was attenuated by 50% in F-11 with constitutively active Akt. The Akt protection was confirmed by DNA fragmentation studies. Both neo-transfected and the c-Akt dominant-negative transfected F-11 cells showed increased ceramide formation (twofold) in response to staurosporine, wortmannin, or LY294002; whereas cells with a constitutively active Akt (Myr-Akt) showed no increase in ceramide when treated with staurosporine, wortmannin, or LY294002. Ceramide was a more potent activator of CPP32 and an inducer of apoptosis when added as the native form (hydroxy- or nonhydroxy-), rather than the more water-soluble C(2)-ceramide. Overexpression of PI3K (p110) and Akt protected cells against ceramide-induced apoptosis, suggesting that Ceramide action is upstream of Akt in these cells and suggesting that Akt might be a target for inhibition by ceramide. Both staurosporine and C(2)-ceramide activated the Jun kinase (JNK) cascade and C(2)-ceramide increased caspase-3 (CPP32) activity in cells expressing wild-type c-Jun, but not dominant-negative (TAM-67) c-Jun. We suggest that this pathway is also involved in apoptosis, consistent with the idea that ceramide has multiple kinase and kinase-modulating targets in the apoptotic pathway of neurons. J. Neurosci. Sci. 57:884-893, 1999.

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Ordering the Cytochrome c–initiated Caspase Cascade: Hierarchical Activation of Caspases-2, -3, -6, -7, -8, and -10 in a Caspase-9–dependent Manner

Cited by 1,745 publications

References 65 publications

Caspase-8/FLICE functions as an executioner caspase in anticancer drug-induced apoptosis

Caspase-8/FLICE functions as an executioner caspase in anticancer drug-induced apoptosis

Retinoid-related molecules require caspase 9 for the effective release of Smac and the rapid induction of apoptosis

Overexpression of Akt (protein kinase B) confers protection against apoptosis and prevents formation of ceramide in response to pro-apoptotic stimuli

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