Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis

Anu Srinivasan, Lyndon M. Foster, Maria Pia Testa, Tõnis Örd, Robert Keane, Dale E. Bredesen, Celik Kayalar

Research output: Contribution to journalArticle

99 Citations (Scopus)

Abstract

The ICE/CED-3 family of proteases has been implicated in playing a fundamental role in programmed cell death, Bcl-2 protein represses a number of apoptotic death programs, but the biochemical mechanism of its action is not known. We investigated the activation of ICE/CED-3 proteases induced by three apoptotic stimuli (staurosporine, ceramide, and serum withdrawal) in the neuronal cell line GT1-7 and in cells overexpressing Bcl-2. Rapid activation of a 17 kDa subunit of an activated member of the ICE/CED-3 family is demonstrated by affinity-labeling GT1-7 extracts from apoptotic controls cells with a biotinylated ICE/CED-3 inhibitor. This activation corresponds to an increased ICE/CED-3-like protease activity in extracts measured by a fluorogenic substrate assay. In a cell free system, these extracts induce apoptotic morphological changes in intact nuclei. All three activities are readily inhibited by treatment of control extracts with ICE/CED-3-like protease inhibitors. Overexpressed Bcl-2 inhibits the activation of the 17 kDa protein, the ICE/CED-3-like protease activity in the fluorogenic assay, and the induction of apoptotic morphological changes in HeLa nuclei in the cell-free system, similar to results obtained with ICE/CED-3 protease inhibitors. At the mRNA level, overexpression of Bcl-2 did not alter expression of five members of the ICE/CED-3 family: CPP32, ICE, Mch 2, Nedd 2, and TX. Overexpression of Bcl-2 prevented the apoptosis induced processing of pro-Nedd 2 to the cleaved form. These data suggest that Bcl-2 participates upstream from the function of ICE/CED-3 proteases and may inhibit apoptosis by preventing the post-translational activation of ICE/CED-3 proteases.

Original languageEnglish
Pages (from-to)5654-5660
Number of pages7
JournalJournal of Neuroscience
Volume16
Issue number18
StatePublished - Sep 15 1996

Fingerprint

Peptide Hydrolases
Apoptosis
Cell-Free System
Protease Inhibitors
Staurosporine
Ceramides
Fluorescent Dyes
Proteins
Cell Death
Cell Line
Messenger RNA
Serum

Keywords

  • apoptosis
  • Bcl-2
  • ceramide
  • ICE/CED-3 proteases
  • neural cells
  • programmed cell death
  • serum withdrawal
  • staurosporine

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Srinivasan, A., Foster, L. M., Testa, M. P., Örd, T., Keane, R., Bredesen, D. E., & Kayalar, C. (1996). Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis. Journal of Neuroscience, 16(18), 5654-5660.

Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis. / Srinivasan, Anu; Foster, Lyndon M.; Testa, Maria Pia; Örd, Tõnis; Keane, Robert; Bredesen, Dale E.; Kayalar, Celik.

In: Journal of Neuroscience, Vol. 16, No. 18, 15.09.1996, p. 5654-5660.

Research output: Contribution to journalArticle

Srinivasan, A, Foster, LM, Testa, MP, Örd, T, Keane, R, Bredesen, DE & Kayalar, C 1996, 'Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis', Journal of Neuroscience, vol. 16, no. 18, pp. 5654-5660.
Srinivasan A, Foster LM, Testa MP, Örd T, Keane R, Bredesen DE et al. Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis. Journal of Neuroscience. 1996 Sep 15;16(18):5654-5660.
Srinivasan, Anu ; Foster, Lyndon M. ; Testa, Maria Pia ; Örd, Tõnis ; Keane, Robert ; Bredesen, Dale E. ; Kayalar, Celik. / Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis. In: Journal of Neuroscience. 1996 ; Vol. 16, No. 18. pp. 5654-5660.
@article{43829fa1ce0a463db3cb8f03d38a893a,
title = "Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis",
abstract = "The ICE/CED-3 family of proteases has been implicated in playing a fundamental role in programmed cell death, Bcl-2 protein represses a number of apoptotic death programs, but the biochemical mechanism of its action is not known. We investigated the activation of ICE/CED-3 proteases induced by three apoptotic stimuli (staurosporine, ceramide, and serum withdrawal) in the neuronal cell line GT1-7 and in cells overexpressing Bcl-2. Rapid activation of a 17 kDa subunit of an activated member of the ICE/CED-3 family is demonstrated by affinity-labeling GT1-7 extracts from apoptotic controls cells with a biotinylated ICE/CED-3 inhibitor. This activation corresponds to an increased ICE/CED-3-like protease activity in extracts measured by a fluorogenic substrate assay. In a cell free system, these extracts induce apoptotic morphological changes in intact nuclei. All three activities are readily inhibited by treatment of control extracts with ICE/CED-3-like protease inhibitors. Overexpressed Bcl-2 inhibits the activation of the 17 kDa protein, the ICE/CED-3-like protease activity in the fluorogenic assay, and the induction of apoptotic morphological changes in HeLa nuclei in the cell-free system, similar to results obtained with ICE/CED-3 protease inhibitors. At the mRNA level, overexpression of Bcl-2 did not alter expression of five members of the ICE/CED-3 family: CPP32, ICE, Mch 2, Nedd 2, and TX. Overexpression of Bcl-2 prevented the apoptosis induced processing of pro-Nedd 2 to the cleaved form. These data suggest that Bcl-2 participates upstream from the function of ICE/CED-3 proteases and may inhibit apoptosis by preventing the post-translational activation of ICE/CED-3 proteases.",
keywords = "apoptosis, Bcl-2, ceramide, ICE/CED-3 proteases, neural cells, programmed cell death, serum withdrawal, staurosporine",
author = "Anu Srinivasan and Foster, {Lyndon M.} and Testa, {Maria Pia} and T{\~o}nis {\"O}rd and Robert Keane and Bredesen, {Dale E.} and Celik Kayalar",
year = "1996",
month = "9",
day = "15",
language = "English",
volume = "16",
pages = "5654--5660",
journal = "Journal of Neuroscience",
issn = "0270-6474",
publisher = "Society for Neuroscience",
number = "18",

}

TY - JOUR

T1 - Bcl-2 expression in neural cells blocks activation of ICE/CED-3 family proteases during apoptosis

AU - Srinivasan, Anu

AU - Foster, Lyndon M.

AU - Testa, Maria Pia

AU - Örd, Tõnis

AU - Keane, Robert

AU - Bredesen, Dale E.

AU - Kayalar, Celik

PY - 1996/9/15

Y1 - 1996/9/15

N2 - The ICE/CED-3 family of proteases has been implicated in playing a fundamental role in programmed cell death, Bcl-2 protein represses a number of apoptotic death programs, but the biochemical mechanism of its action is not known. We investigated the activation of ICE/CED-3 proteases induced by three apoptotic stimuli (staurosporine, ceramide, and serum withdrawal) in the neuronal cell line GT1-7 and in cells overexpressing Bcl-2. Rapid activation of a 17 kDa subunit of an activated member of the ICE/CED-3 family is demonstrated by affinity-labeling GT1-7 extracts from apoptotic controls cells with a biotinylated ICE/CED-3 inhibitor. This activation corresponds to an increased ICE/CED-3-like protease activity in extracts measured by a fluorogenic substrate assay. In a cell free system, these extracts induce apoptotic morphological changes in intact nuclei. All three activities are readily inhibited by treatment of control extracts with ICE/CED-3-like protease inhibitors. Overexpressed Bcl-2 inhibits the activation of the 17 kDa protein, the ICE/CED-3-like protease activity in the fluorogenic assay, and the induction of apoptotic morphological changes in HeLa nuclei in the cell-free system, similar to results obtained with ICE/CED-3 protease inhibitors. At the mRNA level, overexpression of Bcl-2 did not alter expression of five members of the ICE/CED-3 family: CPP32, ICE, Mch 2, Nedd 2, and TX. Overexpression of Bcl-2 prevented the apoptosis induced processing of pro-Nedd 2 to the cleaved form. These data suggest that Bcl-2 participates upstream from the function of ICE/CED-3 proteases and may inhibit apoptosis by preventing the post-translational activation of ICE/CED-3 proteases.

AB - The ICE/CED-3 family of proteases has been implicated in playing a fundamental role in programmed cell death, Bcl-2 protein represses a number of apoptotic death programs, but the biochemical mechanism of its action is not known. We investigated the activation of ICE/CED-3 proteases induced by three apoptotic stimuli (staurosporine, ceramide, and serum withdrawal) in the neuronal cell line GT1-7 and in cells overexpressing Bcl-2. Rapid activation of a 17 kDa subunit of an activated member of the ICE/CED-3 family is demonstrated by affinity-labeling GT1-7 extracts from apoptotic controls cells with a biotinylated ICE/CED-3 inhibitor. This activation corresponds to an increased ICE/CED-3-like protease activity in extracts measured by a fluorogenic substrate assay. In a cell free system, these extracts induce apoptotic morphological changes in intact nuclei. All three activities are readily inhibited by treatment of control extracts with ICE/CED-3-like protease inhibitors. Overexpressed Bcl-2 inhibits the activation of the 17 kDa protein, the ICE/CED-3-like protease activity in the fluorogenic assay, and the induction of apoptotic morphological changes in HeLa nuclei in the cell-free system, similar to results obtained with ICE/CED-3 protease inhibitors. At the mRNA level, overexpression of Bcl-2 did not alter expression of five members of the ICE/CED-3 family: CPP32, ICE, Mch 2, Nedd 2, and TX. Overexpression of Bcl-2 prevented the apoptosis induced processing of pro-Nedd 2 to the cleaved form. These data suggest that Bcl-2 participates upstream from the function of ICE/CED-3 proteases and may inhibit apoptosis by preventing the post-translational activation of ICE/CED-3 proteases.

KW - apoptosis

KW - Bcl-2

KW - ceramide

KW - ICE/CED-3 proteases

KW - neural cells

KW - programmed cell death

KW - serum withdrawal

KW - staurosporine

UR - http://www.scopus.com/inward/record.url?scp=0029816144&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029816144&partnerID=8YFLogxK

M3 - Article

C2 - 8795621

AN - SCOPUS:0029816144

VL - 16

SP - 5654

EP - 5660

JO - Journal of Neuroscience

JF - Journal of Neuroscience

SN - 0270-6474

IS - 18

ER -