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J Biol Chem, Vol. 273, Issue 38, 24884-24890, September 18, 1998
From the cAMP response element-binding protein (CREB) and
activating transcription factor 1 (ATF-1), members of the CREB/ATF
family, have been implicated in cAMP- and calcium-induced
transcriptional activation. We have previously demonstrated that
quenching of CREB-associated proteins in metastatic melanoma cells by a
dominant-negative CREB (KCREB) that is mutated within its DNA-binding
domain decreased their radiation resistance, and their tumorigenic and
metastatic potential in nude mice. As the induction of apoptosis by
diverse exogenous signals is dependent on the elevation of
intracellular Ca2+, the purpose of this study was to
determine the role of CREB and its associated proteins in apoptosis
using KCREB. We used thapsigargin (Tg), which inhibits endoplasmic
reticulum-dependent Ca2+-ATPase and thereby
increases cytosolic Ca2+, to induce apoptosis. MeWo human
melanoma cells were transfected with the KCREB expression vector and
subsequently analyzed for their susceptibility to Tg-induced apoptosis.
Here we demonstrate that expression of KCREB in MeWo cells rendered
them susceptible to Tg-induced apoptosis. Tg treatment induced
phosphorylation of CREB and possibly ATF-1 transcription factors.
Treatment with Tg induced CRE-dependent transcription in
parental cells, whereas this activation was reduced in the
KCREB-transfected cells. In addition, CAT activity driven by the
CRE-dependent promoter was inhibited in parental MeWo cells
cotransfected with increasing concentrations of KCREB in a
dose-dependent manner. We did not observe any changes in
Bcl-2 or Bcl-2-related proteins (Bcl-x, Bax, and Bad) in control or
KCREB-transfected cells before or after treatment with Tg.
Collectively, these data indicate that CREB and its associated proteins
act as survival factors for human melanoma cells, and hence contribute
to the acquisition of the malignant phenotype.
CREB and Its Associated Proteins Act as Survival Factors for
Human Melanoma Cells
,,,
Department of Cell Biology, University of
Texas M. D. Anderson Cancer Center, Houston, Texas 77030 and the
§ Ruttenberg Cancer Center, Mount Sinai School of Medicine,
New York, New York 10029
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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