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(Received for publication, May 30, 1996, and in revised form, August 6, 1996)
From the Departments of In the study presented here, we investigated the
possible interactions between CD95 (Fas/APO-1) and Bcl-2 by studying
the effects of Bcl-2 on the modulation of cellular pathways activated by CD95 using HeLa cells as a model system. We report that stable expression of Bcl-2 in HeLa cells is associated with multiple phenotypic changes. Treatment of HeLa cells with anti-CD95 monoclonal antibody (mAb) resulted in preferential degradation of lamin B compared
with lamins A and C. Significant lamin B degradation was detected as
early as 1 h after anti-CD95 mAb treatment. In contrast, lamins A
and C as well as actin remained unchanged until 4 h after
treatment with anti-CD95 mAb, a time point that correlated with the
period of DNA fragmentation. These results indicate that selective
degradation of lamin B is an early cellular event in response to
activation of the CD95 pathway and that it precedes DNA fragmentation.
Overexpression of Bcl-2 resulted in prevention of lamin B degradation
and DNA fragmentation into oligonucleosome fragments in response to the
apoptotic signal by anti-CD95 mAb. In addition, in Bcl-2-overexpressing
cells that were protected against apoptosis, anti-CD95 mAb-induced
cleavage of poly(ADP-ribose) polymerase was completely blocked.
Overexpression of Bcl-2 also resulted in restoration of the
CD95-mediated signaling pathway involving activation of the
transcription factor NF-
Volume 271, Number 48,
Issue of November 29, 1996
pp. 30354-30359
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
B
Signaling Pathway
§
,§
,
,§
Medicine,
§ Cellular and Molecular Physiology, and ¶ Microbiology
and Immunology, Pennsylvania State University College of Medicine,
Hershey, Pennsylvania 17033 and the 
Division of Oncology
Research, Mayo Medical School, Rochester, Minnesota 55905
B (p50/RelA). These findings suggest that
Bcl-2 prevents apoptosis in part by preventing the degradation of major
nuclear polypeptides such as lamin B and poly(ADP-ribose) polymerase.
In addition, our results demonstrate that CD95-mediated signaling
involves activation of NF-B (p50/RelA).
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