Breast cancer cell proliferation is inhibited by BAD

Regulation of cyclin D1

Romaine Fernando, James S. Foster, Amber Bible, Anders Ström, Richard G. Pestell, Mahadev Rao, Arnold Saxton, Joon Baek Seung, Kiyoshi Yamaguchi, Robert Donnell, Maria Cekanova, Jay Wimalasena

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Recent investigations suggest that functions of the proapoptotic BCL2 family members, including BAD, are not limited to regulation of apoptosis. Here we demonstrate that BAD inhibits G1 to S phase transition in MCF7 breast cancer cells independent of apoptosis. BAD overexpression inhibited G1 transit and cell growth as well as cyclin D1 expression. Inhibition of cyclin D1 expression was mediated through inhibition of transcription activated by AP1. Chromatin immunoprecipitation assays indicated that BAD is localized at the 12-O-tetradecanoylphorbol-13-acetate-response element (TRE) and cAMP-response element (CRE) in the cyclin D1 promoter. This was shown to reflect direct binding interactions of BAD with c-Jun, and this interaction inhibited the activity of AP1 complexes at TRE. BAD did not interact with phosphorylated forms of c-Jun. Our data suggest that inhibitory TRE/CRE-c-Jun-BAD complexes are present at the cyclin D1 promoter in quiescent cells. Estrogen stimulation displaced BAD from TRE/CRE elements in MCF7 cells, whereas BAD overexpression inhibited estrogen-induced cyclin D1 synthesis and cell proliferation. Inhibition of endogenous BAD in MCF7 cells markedly increased the proliferative fraction and DNA synthesis, activated Cdks, and increased cyclin D1 protein levels. This action of BAD required serine residues Ser75 and Ser99. Both phosphorylated and unphosphorylated forms of BAD localized to the nuclei of human breast epithelial cells. Thus, we demonstrate a novel role for BAD in cell cycle regulation dependent upon its phosphorylation state and independent of the BAD/BCL2 interaction and apoptosis.

Original languageEnglish
Pages (from-to)28864-28873
Number of pages10
JournalJournal of Biological Chemistry
Volume282
Issue number39
DOIs
Publication statusPublished - 28-09-2007

Fingerprint

Cyclin D1
Cell proliferation
Response Elements
Cell Proliferation
Breast Neoplasms
Tetradecanoylphorbol Acetate
Acetates
MCF-7 Cells
Apoptosis
Estrogens
Cells
Phosphorylation
Chromatin Immunoprecipitation
Phase Transition
Cell growth
Transcription
S Phase
Serine
Chromatin
Assays

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Fernando, R., Foster, J. S., Bible, A., Ström, A., Pestell, R. G., Rao, M., ... Wimalasena, J. (2007). Breast cancer cell proliferation is inhibited by BAD: Regulation of cyclin D1. Journal of Biological Chemistry, 282(39), 28864-28873. https://doi.org/10.1074/jbc.M700785200
Fernando, Romaine ; Foster, James S. ; Bible, Amber ; Ström, Anders ; Pestell, Richard G. ; Rao, Mahadev ; Saxton, Arnold ; Seung, Joon Baek ; Yamaguchi, Kiyoshi ; Donnell, Robert ; Cekanova, Maria ; Wimalasena, Jay. / Breast cancer cell proliferation is inhibited by BAD : Regulation of cyclin D1. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 39. pp. 28864-28873.
@article{32369c3de2f64ace86cf8b95155e8af0,
title = "Breast cancer cell proliferation is inhibited by BAD: Regulation of cyclin D1",
abstract = "Recent investigations suggest that functions of the proapoptotic BCL2 family members, including BAD, are not limited to regulation of apoptosis. Here we demonstrate that BAD inhibits G1 to S phase transition in MCF7 breast cancer cells independent of apoptosis. BAD overexpression inhibited G1 transit and cell growth as well as cyclin D1 expression. Inhibition of cyclin D1 expression was mediated through inhibition of transcription activated by AP1. Chromatin immunoprecipitation assays indicated that BAD is localized at the 12-O-tetradecanoylphorbol-13-acetate-response element (TRE) and cAMP-response element (CRE) in the cyclin D1 promoter. This was shown to reflect direct binding interactions of BAD with c-Jun, and this interaction inhibited the activity of AP1 complexes at TRE. BAD did not interact with phosphorylated forms of c-Jun. Our data suggest that inhibitory TRE/CRE-c-Jun-BAD complexes are present at the cyclin D1 promoter in quiescent cells. Estrogen stimulation displaced BAD from TRE/CRE elements in MCF7 cells, whereas BAD overexpression inhibited estrogen-induced cyclin D1 synthesis and cell proliferation. Inhibition of endogenous BAD in MCF7 cells markedly increased the proliferative fraction and DNA synthesis, activated Cdks, and increased cyclin D1 protein levels. This action of BAD required serine residues Ser75 and Ser99. Both phosphorylated and unphosphorylated forms of BAD localized to the nuclei of human breast epithelial cells. Thus, we demonstrate a novel role for BAD in cell cycle regulation dependent upon its phosphorylation state and independent of the BAD/BCL2 interaction and apoptosis.",
author = "Romaine Fernando and Foster, {James S.} and Amber Bible and Anders Str{\"o}m and Pestell, {Richard G.} and Mahadev Rao and Arnold Saxton and Seung, {Joon Baek} and Kiyoshi Yamaguchi and Robert Donnell and Maria Cekanova and Jay Wimalasena",
year = "2007",
month = "9",
day = "28",
doi = "10.1074/jbc.M700785200",
language = "English",
volume = "282",
pages = "28864--28873",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "39",

}

Fernando, R, Foster, JS, Bible, A, Ström, A, Pestell, RG, Rao, M, Saxton, A, Seung, JB, Yamaguchi, K, Donnell, R, Cekanova, M & Wimalasena, J 2007, 'Breast cancer cell proliferation is inhibited by BAD: Regulation of cyclin D1', Journal of Biological Chemistry, vol. 282, no. 39, pp. 28864-28873. https://doi.org/10.1074/jbc.M700785200

Breast cancer cell proliferation is inhibited by BAD : Regulation of cyclin D1. / Fernando, Romaine; Foster, James S.; Bible, Amber; Ström, Anders; Pestell, Richard G.; Rao, Mahadev; Saxton, Arnold; Seung, Joon Baek; Yamaguchi, Kiyoshi; Donnell, Robert; Cekanova, Maria; Wimalasena, Jay.

In: Journal of Biological Chemistry, Vol. 282, No. 39, 28.09.2007, p. 28864-28873.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Breast cancer cell proliferation is inhibited by BAD

T2 - Regulation of cyclin D1

AU - Fernando, Romaine

AU - Foster, James S.

AU - Bible, Amber

AU - Ström, Anders

AU - Pestell, Richard G.

AU - Rao, Mahadev

AU - Saxton, Arnold

AU - Seung, Joon Baek

AU - Yamaguchi, Kiyoshi

AU - Donnell, Robert

AU - Cekanova, Maria

AU - Wimalasena, Jay

PY - 2007/9/28

Y1 - 2007/9/28

N2 - Recent investigations suggest that functions of the proapoptotic BCL2 family members, including BAD, are not limited to regulation of apoptosis. Here we demonstrate that BAD inhibits G1 to S phase transition in MCF7 breast cancer cells independent of apoptosis. BAD overexpression inhibited G1 transit and cell growth as well as cyclin D1 expression. Inhibition of cyclin D1 expression was mediated through inhibition of transcription activated by AP1. Chromatin immunoprecipitation assays indicated that BAD is localized at the 12-O-tetradecanoylphorbol-13-acetate-response element (TRE) and cAMP-response element (CRE) in the cyclin D1 promoter. This was shown to reflect direct binding interactions of BAD with c-Jun, and this interaction inhibited the activity of AP1 complexes at TRE. BAD did not interact with phosphorylated forms of c-Jun. Our data suggest that inhibitory TRE/CRE-c-Jun-BAD complexes are present at the cyclin D1 promoter in quiescent cells. Estrogen stimulation displaced BAD from TRE/CRE elements in MCF7 cells, whereas BAD overexpression inhibited estrogen-induced cyclin D1 synthesis and cell proliferation. Inhibition of endogenous BAD in MCF7 cells markedly increased the proliferative fraction and DNA synthesis, activated Cdks, and increased cyclin D1 protein levels. This action of BAD required serine residues Ser75 and Ser99. Both phosphorylated and unphosphorylated forms of BAD localized to the nuclei of human breast epithelial cells. Thus, we demonstrate a novel role for BAD in cell cycle regulation dependent upon its phosphorylation state and independent of the BAD/BCL2 interaction and apoptosis.

AB - Recent investigations suggest that functions of the proapoptotic BCL2 family members, including BAD, are not limited to regulation of apoptosis. Here we demonstrate that BAD inhibits G1 to S phase transition in MCF7 breast cancer cells independent of apoptosis. BAD overexpression inhibited G1 transit and cell growth as well as cyclin D1 expression. Inhibition of cyclin D1 expression was mediated through inhibition of transcription activated by AP1. Chromatin immunoprecipitation assays indicated that BAD is localized at the 12-O-tetradecanoylphorbol-13-acetate-response element (TRE) and cAMP-response element (CRE) in the cyclin D1 promoter. This was shown to reflect direct binding interactions of BAD with c-Jun, and this interaction inhibited the activity of AP1 complexes at TRE. BAD did not interact with phosphorylated forms of c-Jun. Our data suggest that inhibitory TRE/CRE-c-Jun-BAD complexes are present at the cyclin D1 promoter in quiescent cells. Estrogen stimulation displaced BAD from TRE/CRE elements in MCF7 cells, whereas BAD overexpression inhibited estrogen-induced cyclin D1 synthesis and cell proliferation. Inhibition of endogenous BAD in MCF7 cells markedly increased the proliferative fraction and DNA synthesis, activated Cdks, and increased cyclin D1 protein levels. This action of BAD required serine residues Ser75 and Ser99. Both phosphorylated and unphosphorylated forms of BAD localized to the nuclei of human breast epithelial cells. Thus, we demonstrate a novel role for BAD in cell cycle regulation dependent upon its phosphorylation state and independent of the BAD/BCL2 interaction and apoptosis.

UR - http://www.scopus.com/inward/record.url?scp=35349029694&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=35349029694&partnerID=8YFLogxK

U2 - 10.1074/jbc.M700785200

DO - 10.1074/jbc.M700785200

M3 - Article

VL - 282

SP - 28864

EP - 28873

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 39

ER -