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J. Biol. Chem., Vol. 275, Issue 21, 15691-15700, May 26, 2000
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From the Department of Biochemistry and Molecular Biology,
Georgetown University Medical Center, Washington, D. C. 20007
Exposure to anti-Fas antibody in Jurkat cells
(type II cells), which are characterized by a weak caspase-8 activation
at the death-inducing signaling complex (DISC), induced a biphasic
increase in ceramide levels. The early generation of ceramide preceded transient activation of acidic ceramidase and subsequent production of
sphingosine, followed by cytochrome c release, activation
of caspases-2, -3, -6, -7, -8, and -9, Bid cleavage, and a later sustained ceramide accumulation. The caspase inhibitor
benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone inhibited early
increases of ceramide and sphingosine, whereas overexpression of
Bcl-xL had no effect, and both prevented the later
sustained ceramide accumulation. Exogenous sphingosine, as well as
cell-permeable C2-ceramide, induced cytochrome
c release from mitochondria in a caspase-independent
fashion leading to activation of caspase-9 and executioner caspases
and, surprisingly, activation of the initiator caspase-8 and processing
of its substrate Bid. These effects were also completely abolished by
Bcl-xL overexpression. Our results suggest that sphingosine
might also be involved in the mitochondria-mediated pathway of
Fas-induced cell death in type II cells.
Involvement of Sphingosine in Mitochondria-dependent
Fas-induced Apoptosis of Type II Jurkat T Cells*
,
*
This work was supported by Grant GM3880 from the National
Institutes of Health (to S. S.).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.
Present address: Unité Inserm 466, CHU Rangueil, 1 Ave. Jean
Poulhès, 31403 Toulouse Cedex 4, France.
§
To whom correspondence should be addressed: Dept. of Biochemistry
and Molecular Biology, Georgetown University Medical Center, 353 Basic
Science Bldg., 3900 Reservoir Rd. NW, Washington, D. C. 20007. Tel.:
202-687-1432; Fax: 202-687-0260; E-mail:
spiegel@bc.georgetown.edu.
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