CREB and its associated proteins act as survival factors for human melanoma cells

Didier Jean, Matthew Harbison, David McConkey, Zeèv Ronai, Menashe Bar-Eli

Research output: Contribution to journalArticle

Abstract

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 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.

Original languageEnglish (US)
Pages (from-to)24884-24890
Number of pages7
JournalJournal of Biological Chemistry
Volume273
Issue number38
DOIs
StatePublished - Sep 18 1998
Externally publishedYes

Fingerprint

Cyclic AMP Response Element-Binding Protein
Thapsigargin
Melanoma
Survival
Apoptosis
Activating Transcription Factor 1
Proteins
Chemical activation
Phosphorylation
Calcium-Transporting ATPases
Transcription
Quenching
Nude Mice
Endoplasmic Reticulum
Transcriptional Activation
Transcription Factors
Calcium
Radiation
Phenotype
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

CREB and its associated proteins act as survival factors for human melanoma cells. / Jean, Didier; Harbison, Matthew; McConkey, David; Ronai, Zeèv; Bar-Eli, Menashe.

In: Journal of Biological Chemistry, Vol. 273, No. 38, 18.09.1998, p. 24884-24890.

Research output: Contribution to journalArticle

Jean, Didier ; Harbison, Matthew ; McConkey, David ; Ronai, Zeèv ; Bar-Eli, Menashe. / CREB and its associated proteins act as survival factors for human melanoma cells. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 38. pp. 24884-24890.
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