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J Biol Chem, Vol. 273, Issue 28, 17477-17482, July 10, 1998
Signal Transduction
§ and
From the Reactive oxygen species (ROS) have been
implicated in the induction of apoptosis by tumor necrosis factor-
Fred Hutchinson Cancer Research Center,
A2-025, Seattle, Washington 98109 and the § Division of
Neuroscience, Children's Hospital, and Department of Neurobiology,
Harvard Medical School, Boston, Massachusetts 02115
(TNF
) and other cytotoxic insults, although the molecule(s)
regulated by ROS in TNF
signaling have not been identified.
Apoptosis signal-regulating kinase 1 (ASK1) is a member of the
mitogen-activated protein kinase kinase kinase (MAPKKK) superfamily
that has been shown to be activated during TNF
-induced apoptosis.
ASK1 increases apoptosis when overexpressed, but the mechanism of ASK1
activation and the mechanisms of ASK1-induced apoptosis are unclear. We
now report that hydrogen peroxide induces the activation of ASK1 in 293 cells. TNF
-induced activation of ASK1 was inhibited by antioxidants.
Hydrogen peroxide-induced apoptosis was markedly enhanced by the
expression of ASK1. These results suggest that TNF
-induced
activation of ASK1 is mediated by ROS. We also examined how ASK1
activity is regulated by ROS. We found that ASK1 formed dimers or
higher order oligomers in 293 cells. TNF
or hydrogen peroxide
treatment increased the dimeric form of ASK1, and pretreatment with
N-acetylcysteine decreased it. Furthermore, synthetic
dimerization of an ASK1-gyrase B fusion protein by coumermycin resulted
in substantial activation of ASK1, suggesting that dimerization of ASK1
is sufficient for its activation. These results taken together suggest
that TNF
causes ASK1 activation via ROS-mediated dimerization of
ASK1.
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