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J Biol Chem, Vol. 273, Issue 7, 4027-4034, February 13, 1998
in Rat
Mesangial Cells
From the Department of Biochemistry and Biophysics, School of
Medicine, University of Pennsylvania,
Philadelphia, Pennsylvania 19104
Rat mesangial cells are normally resistant to
tumor necrosis factor-
(TNF-)-induced apoptosis. In this report
we show that the cells can be made susceptible to the apoptotic effect
of TNF- when pretreated with actinomycin D, cycloheximide, or
vanadate. c-Jun N-terminal protein kinase (JNK) has been thought to
mediate apoptotic processes elicited by some stimuli, but its
involvement in TNF--induced apoptosis has been controversial. JNK
activation was investigated under conditions where the mesangial cells
were either resistant or susceptible to TNF--induced apoptosis.
TNF- alone stimulated a single transient JNK activity peak. However, when the cells were pretreated with actinomycin D or cycloheximide, TNF- stimulated a second sustained JNK activity peak. When the cells
were pretreated with the phosphatase inhibitor vanadate, TNF--induced JNK activation was greatly prolonged. In all three cases, a sustained JNK activation was associated with the initiation of
apoptosis. Our data suggest that a sustained activation of JNK induced
by these reagents may be associated with blocking the expression of a
phosphatase that inactivates JNK. Further studies reveal that the
expression of mitogen-activated protein kinase phosphatase-1 (MKP-1)
was induced by TNF-, indicating that MKP-1 may be involved in
protecting the cells from apoptosis by preventing a prolonged
activation of JNK under normal conditions. Additional studies
showed that extracellular signal-regulated protein kinase activation
stimulated by TNF- was unlikely to contribute to the resistance of
mesangial cells to TNF- cytotoxicity.
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