Different Cleavage Pattern for Poly(ADP-Ribose) Polymerase during Necrosis and Apoptosis in HL-60 Cells

Shah, Girish M.; Shah, Rashmi G.; Poirier, Guy G.

doi:10.1006/bbrc.1996.1889

Cited by 155 publications

(96 citation statements)

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“…Interestingly, the levels of µ-and m-calpain increased from DIV10 ( Figure 3B). These findings agree with the observed decrease in Bax and with the early cleavage of PARP, which preceded caspase-3 processing, because Bax, caspase-3, and PARP are proteolytically processed by calpains (Kobayashi et al, 1990;Shah et al, 1996;Wood et al, 1998;McGinnis et al, 1999b). Thus, we examined the levels of µ-and m-calpain.…”

Section: Patterns Of Apoptotic Molecules During Cultivationsupporting

confidence: 86%

Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Kim

Oh²,

Park³

et al. 2007

Exp Mol Med

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Primary neuronal culture is a powerful tool to study neuronal development, aging, and degeneration. However, cultured neurons show signs of cell death after 2 or 3 weeks. Although the mechanism underlying this phenomenon has not been elucidated, several preventive methods have been identified. Here we show that the neuronal loss in primary cortical culture involves calpain activation and subsequent neuronal cell death. Neuronal loss during cultivation showed destruction of neurites and synapses, and a decrease in neuron numbers. µ-Calpain and m-calpain were initially activated and accumulated by increased RNA expression. This neuronal death exhibited neurodegenerative features, such as conversion of p35 to p25, which is important in the developmental process and in the pathogenesis of Alzheimer's disease. But, postnatal and aged rat cortex did not show calpain activation and prolonged processing of p35 to p25, in contrast to the long-term culture of cortical neurons. In addition, the inhibition of calpains by ALLM or ALLN blocked the conversion of p35 to p25, indicating that the calpain activity is essential for the neurodegenerative features of cell death. Taken together, this study shows that the neuronal loss in primary cortical cultures involves neurodegeneration-like cell death through the activation of calpains and the subsequent processing of p35 to p25, but not developmental apoptosis or aging. Our results suggest that the long term primary culture of cortical neurons represent a valuable model of neurodegeneration, such as Alzheimer's disease.

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Section: Patterns Of Apoptotic Molecules During Cultivationsupporting

confidence: 86%

Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Kim

Oh²,

Park³

et al. 2007

Exp Mol Med

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“…The 50 kDa fragment was designated as a major necrotic fragment. [38][39][40] From these results, we confirmed that the death of HepG2 cells induced by CCl 4 is predominantly due to necrosis.…”

Section: Necrosis In Hepg2 Induced By CCLmentioning

confidence: 53%

“…Detection of a 50 kDa fragment derived from PARP-1 in CCl 4 -treated cells is strong evidence for necrosis, because the apoptotic PARP-1 fragment of 85 kDa induced with TNFa/CHX is distinct from the 50 kDa fragment 38,39 (Figure 1d). These results clearly indicated that a majority of HepG2 cells exposed to CCl 4 died in a necrotic manner, with a good agreement with previous results.…”

Section: Discussionmentioning

confidence: 99%

Zonal necrosis prevented by transduction of the artificial anti-death FNK protein

et al. 2005

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Protection of cells from necrosis would be important for many medical applications. Here, we show protein transduction domain (PTD)-FNK therapeutics based on protein transduction to prevent necrosis and acute hepatic injury with zonal death induced by carbon tetrachloride (CCl 4 ). PTD-FNK is a fusion protein comprising the HIV/Tat PTD and FNK, a gain-offunction mutant of anti-apoptotic Bcl-x L . PTD-FNK protected hepatoma HepG2 from necrotic death induced by CCl 4 , and additionally, increased the apoptotic population among cells treated with CCl 4 . A concomitant treatment with a pancaspase inhibitor Z-VAD-FMK (N-benzyloxycarbonyl-Val-AlaAsp-fluoromethylketone), which alone could not prevent the necrosis, protected these cells from the apoptosis. When preinjected intraperitoneally, PTD-FNK markedly reduced zonal liver necrosis caused by CCl 4 . Moreover, injection of PTD-FNK accompanied by Z-VAD-FMK suppressed necrotic injury even after CCl 4 administration. These results suggest that PTD-FNK has great potential for clinical applications to prevent cell death, whether from apoptosis or necrosis, and organ failure.

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“…As a positive control, we used PARP1 which is cleaved by Calpain to produce several fragments about 35, 45, 50 and 89 kDa in size, in contrast to the single 89 kDa band produced by caspase cleavage of PARP1. 21 Treatment of HL60 cells for 90 min with Ionomycin caused almost complete cleavage of PARP1 into small molecular weight fragments, whereas the levels of Claspin remained practically the same (Figure 4f). Treatment of HL60 cells with Ionomycin together with Calpain inhibitor II (ALLM) inhibited the degradation of PARP1 and caused a striking increase in the levels of Claspin, that was also seen when Claspin was treated by ALLM alone (Figure 4f).…”

Section: Proteasomal Degradation Of Claspin During Apoptosismentioning

confidence: 92%

Cleavage and degradation of Claspin during apoptosis by caspases and the proteasome

et al. 2007

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Apoptosis plays a crucial role in development and tissue homeostasis. Some key survival pathways, such as DNA damage checkpoints and DNA repair, have been described to be inactivated during apoptosis. Here, we describe the processing of the human checkpoint protein Claspin during apoptosis. We observed cleavage of Claspin into multiple fragments in vivo. In vitro cleavage with caspases 3 and 7 of various fragments of the protein, revealed cut sites near the N-and C-termini of the protein.Using mass spectrometry, we identified a novel caspase cleavage site in Claspin at Asp25. Importantly, in addition to cleavage by caspases, we observed a proteasome-dependent degradation of Claspin under apoptotic conditions, resulting in a reduction of the levels of both full-length Claspin and its cleavage products. This degradation was not dependent upon the DSGxxS phosphodegron motif required for SCF b-TrCP -mediated ubiquitination of Claspin. Finally, downregulation of Claspin protein levels by short interfering RNA resulted in an increase in apoptotic induction both in the presence and absence of DNA damage. We conclude that Claspin has antiapoptotic activity and is degraded by two different pathways during apoptosis. The resulting disappearance of Claspin from the cells further promotes apoptosis.

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Different Cleavage Pattern for Poly(ADP-Ribose) Polymerase during Necrosis and Apoptosis in HL-60 Cells

Cited by 155 publications

References 0 publications

Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Zonal necrosis prevented by transduction of the artificial anti-death FNK protein

Cleavage and degradation of Claspin during apoptosis by caspases and the proteasome

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