Ordering the Cell Death Pathway

doi:10.1074/jbc.271.36.21898

Ordering the Cell Death Pathway

DIFFERENTIAL EFFECTS OF BCL2, AN INTERLEUKIN-1-CONVERTING ENZYME FAMILY PROTEASE INHIBITOR, AND OTHER SURVIVAL AGENTS ON JNK ACTIVATION IN SERUM/NERVE GROWTH FACTOR-DEPRIVED PC12 CELLS*

From the Department of Pathology and Center for Neurobiology and Behavior, Columbia University, College of Physicians and Surgeons, New York, New York 10032

^‡ Recipient of an Aaron Diamond Foundation Fellowship. To whom correspondence should be addressed. Tel.: 212-305-6370; Fax: 212-305-5498.

Abstract

Previous studies indicate that activation of c-Jun kinase (JNK) is necessary for apoptosis of trophic factor-deprived PC12 cells and that death in this system is suppressed by multiple agents, including BCL2, inhibitors of the interleukin-1-converting enzyme (ICE) family of proteases, blockers of transcription, and a variety of small molecules with differing modes of action. Here, we determine the order in which these agents block apoptosis relative to JNK activation. Overexpression of BCL2 promotes PC12 cell survival and blocks JNK activation caused by trophic factor withdrawal. Similarly, the survival-promoting agents aurintricarboxylic acid, N-acetylcysteine, the nitric oxide generator diethylenetriamine nitric oxide, 8-bromo-cGMP, and 8-(4-chlorophenylthio)-cAMP act upstream to inhibit JNK activation. In contrast, zVAD-fluoromethylketone (a permeant ICE family inhibitor), actinomycin D, and the G₁/S cell cycle inhibitor deferoxamine, all promote survival after trophic factor withdrawal, but do not affect JNK activation. These findings are consistent with the presence of an ordered cell death pathway triggered by trophic factor deprivation in which 1) BCL2 and a number of survival-promoting agents act upstream of JNK, 2) ICE family protease actions, regulated genes required for cell death, and certain cell cycle blockers lie either downstream of JNK or on independent pathways required for apoptotic death.

Footnotes

↵^§ Supported in part by an National Research Service Award from the NIH-NUNDS.
↵* This work was supported in part by grants from the NIH-NINDS, March of Dimes, and Amyotrophic Lateral Sclerosis Foundation and Aaron Diamond Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵¹ The abbreviations used are:

ICE

interleukin-1-converting enzyme

JNK

Jun kinase

NGF

nerve growth factor

ATA

aurintricarboxylic acid

PKA

cyclic AMP-dependent protein kinase

NAC

N-acetylcysteine

GST

glutathione S-transferase

fmk

fluoromethylketone

DEPA·NO

diethylenetriamine nitric oxide

CPT-cAMP

8-(4-chlorophenylthio)-cAMP

DFX

deferoxamine
↵²Virdee, K., and Tolkovsky, A. M. (1995) Cold Spring Harbor Programmed Cell Death, September 20-24, Cold Spring Harbor, NY.
- Received February 5, 1996.
- Revision received May 16, 1996.