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J Biol Chem, Vol. 275, Issue 2, 1233-1240, January 14, 2000
From the Accelerated apoptosis is one mechanism proposed
for the loss of CD4+ T-lymphocytes in human immunodeficiency virus type
1 (HIV-1) infection. The HIV-1 envelope glycoprotein, gp160, contains two C-terminal calmodulin-binding domains. Expression of
gp160 in Jurkat T-cells results in increased sensitivity to FAS-
and ceramide-mediated apoptosis. The pro-apoptotic effect of gp160 expression is blocked by two calmodulin antagonists, tamoxifen and trifluoperazine. This enhanced apoptosis in response to FAS antibody or C2-ceramide is associated with activation
of caspase 3, a critical mediator of apoptosis. A point mutation in the
C-terminal calmodulin-binding domain of gp160 (alanine 835 to
tryptophan, A835W) eliminates gp160-dependent enhanced
FAS-mediated apoptosis in transiently transfected cells, as well as
in vitro calmodulin binding to a peptide corresponding to
the C-terminal calmodulin-binding domain of gp160. Stable Tet-off
Jurkat cell lines were developed that inducibly express wild type gp160
or gp160A835W. Increasing expression of wild type gp160, but not
gp160A835W, correlates with increased calmodulin levels, increased
apoptosis, and caspase 3 activation in response to anti-FAS treatment.
The data indicate that gp160-enhanced apoptosis is dependent upon
calmodulin up-regulation, involves the activation of caspase 3, and
requires calmodulin binding to the C-terminal binding domain of gp160.
Requirement of Calmodulin Binding by HIV-1 gp160 for Enhanced
FAS-mediated Apoptosis*
,,,,, and§¶
Department of Pathology, University of
Alabama at Birmingham, Birmingham, Alabama 35294 and the
§ Veterans Administration Medical Center,
Birmingham, Alabama 35233
*
This work was supported by NCI Grant CA/72823 (to
J. M. M.) from the National Institutes of Health and a Veterans
Administration Merit Review (to J. M. M.).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.
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