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(Received for publication, February 5, 1996, and in revised form, May 16, 1996)
From the Department of Pathology and Center for Neurobiology and
Behavior, Columbia University, College of Physicians and Surgeons,
New York, New York 10032
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.
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