p53 Modulates Notch Signaling in MCF-7 Breast Cancer Cells by Associating With the Notch Transcriptional Complex Via MAML1

Jieun Yun, Ingrid Espinoza, Antonio Pannuti, Damian Romero, Luis Martinez, Mary Caskey, Adina Stanculescu, Maurizio Bocchetta, Paola Rizzo, Vimla Band, Hamid Band, Hwan Mook Kim, Song Kyu Park, Keon Wook Kang, Maria Laura Avantaggiati, Christian R. Gomez, Todd Golde, Barbara Osborne, Lucio Miele

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

p53 and Notch-1 play important roles in breast cancer biology. Notch-1 inhibits p53 activity in cervical and breast cancer cells. Conversely, p53 inhibits Notch activity in T-cells but stimulates it in human keratinocytes. Notch co-activator MAML1 binds p53 and functions as a p53 co-activator. We studied the regulation of Notch signaling by p53 in MCF-7 cells and normal human mammary epithelial cells (HMEC). Results show that overexpression of p53 or activation of endogenous p53 with Nutlin-3 inhibits Notch-dependent transcriptional activity and Notch target expression in a dose-dependent manner. This effect could be partially rescued by transfection of MAML1 but not p300. Standard and quantitative co-immunoprecipitation experiments readily detected a complex containing p53 and Notch-1 in MCF-7 cells. Formation of this complex was inhibited by dominant negative MAML1 (DN-MAML1) and stimulated by wild-type MAML1. Standard and quantitative far-Western experiments showed a complex including p53, Notch-1, and MAML1. Chromatin immunoprecipitation (ChIP) experiments showed that p53 can associate with Notch-dependent HEY1 promoter and this association is inhibited by DN-MAML1 and stimulated by wild-type MAML1. Our data support a model in which p53 associates with the Notch transcriptional complex (NTC) in a MAML1-dependent fashion, most likely through a p53-MAML1 interaction. In our cellular models, the effect of this association is to inhibit Notch-dependent transcription. Our data suggest that p53-null breast cancers may lack this Notch-modulatory mechanism, and that therapeutic strategies that activate wild-type p53 can indirectly cause inhibition of Notch transcriptional activity.

Original languageEnglish (US)
Pages (from-to)3115-3127
Number of pages13
JournalJournal of Cellular Physiology
Volume230
Issue number12
DOIs
StatePublished - Dec 1 2015

Fingerprint

MCF-7 Cells
Cells
Breast Neoplasms
Association reactions
T-cells
Chromatin Immunoprecipitation
Experiments
Transcription
Keratinocytes
Immunoprecipitation
Uterine Cervical Neoplasms
Chromatin
Transfection
Breast
Epithelial Cells
Chemical activation
T-Lymphocytes
Therapeutics
nutlin 3

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

p53 Modulates Notch Signaling in MCF-7 Breast Cancer Cells by Associating With the Notch Transcriptional Complex Via MAML1. / Yun, Jieun; Espinoza, Ingrid; Pannuti, Antonio; Romero, Damian; Martinez, Luis; Caskey, Mary; Stanculescu, Adina; Bocchetta, Maurizio; Rizzo, Paola; Band, Vimla; Band, Hamid; Kim, Hwan Mook; Park, Song Kyu; Kang, Keon Wook; Avantaggiati, Maria Laura; Gomez, Christian R.; Golde, Todd; Osborne, Barbara; Miele, Lucio.

In: Journal of Cellular Physiology, Vol. 230, No. 12, 01.12.2015, p. 3115-3127.

Research output: Contribution to journalArticle

Yun, J, Espinoza, I, Pannuti, A, Romero, D, Martinez, L, Caskey, M, Stanculescu, A, Bocchetta, M, Rizzo, P, Band, V, Band, H, Kim, HM, Park, SK, Kang, KW, Avantaggiati, ML, Gomez, CR, Golde, T, Osborne, B & Miele, L 2015, 'p53 Modulates Notch Signaling in MCF-7 Breast Cancer Cells by Associating With the Notch Transcriptional Complex Via MAML1', Journal of Cellular Physiology, vol. 230, no. 12, pp. 3115-3127. https://doi.org/10.1002/jcp.25052
Yun, Jieun ; Espinoza, Ingrid ; Pannuti, Antonio ; Romero, Damian ; Martinez, Luis ; Caskey, Mary ; Stanculescu, Adina ; Bocchetta, Maurizio ; Rizzo, Paola ; Band, Vimla ; Band, Hamid ; Kim, Hwan Mook ; Park, Song Kyu ; Kang, Keon Wook ; Avantaggiati, Maria Laura ; Gomez, Christian R. ; Golde, Todd ; Osborne, Barbara ; Miele, Lucio. / p53 Modulates Notch Signaling in MCF-7 Breast Cancer Cells by Associating With the Notch Transcriptional Complex Via MAML1. In: Journal of Cellular Physiology. 2015 ; Vol. 230, No. 12. pp. 3115-3127.
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abstract = "p53 and Notch-1 play important roles in breast cancer biology. Notch-1 inhibits p53 activity in cervical and breast cancer cells. Conversely, p53 inhibits Notch activity in T-cells but stimulates it in human keratinocytes. Notch co-activator MAML1 binds p53 and functions as a p53 co-activator. We studied the regulation of Notch signaling by p53 in MCF-7 cells and normal human mammary epithelial cells (HMEC). Results show that overexpression of p53 or activation of endogenous p53 with Nutlin-3 inhibits Notch-dependent transcriptional activity and Notch target expression in a dose-dependent manner. This effect could be partially rescued by transfection of MAML1 but not p300. Standard and quantitative co-immunoprecipitation experiments readily detected a complex containing p53 and Notch-1 in MCF-7 cells. Formation of this complex was inhibited by dominant negative MAML1 (DN-MAML1) and stimulated by wild-type MAML1. Standard and quantitative far-Western experiments showed a complex including p53, Notch-1, and MAML1. Chromatin immunoprecipitation (ChIP) experiments showed that p53 can associate with Notch-dependent HEY1 promoter and this association is inhibited by DN-MAML1 and stimulated by wild-type MAML1. Our data support a model in which p53 associates with the Notch transcriptional complex (NTC) in a MAML1-dependent fashion, most likely through a p53-MAML1 interaction. In our cellular models, the effect of this association is to inhibit Notch-dependent transcription. Our data suggest that p53-null breast cancers may lack this Notch-modulatory mechanism, and that therapeutic strategies that activate wild-type p53 can indirectly cause inhibition of Notch transcriptional activity.",
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T1 - p53 Modulates Notch Signaling in MCF-7 Breast Cancer Cells by Associating With the Notch Transcriptional Complex Via MAML1

AU - Yun, Jieun

AU - Espinoza, Ingrid

AU - Pannuti, Antonio

AU - Romero, Damian

AU - Martinez, Luis

AU - Caskey, Mary

AU - Stanculescu, Adina

AU - Bocchetta, Maurizio

AU - Rizzo, Paola

AU - Band, Vimla

AU - Band, Hamid

AU - Kim, Hwan Mook

AU - Park, Song Kyu

AU - Kang, Keon Wook

AU - Avantaggiati, Maria Laura

AU - Gomez, Christian R.

AU - Golde, Todd

AU - Osborne, Barbara

AU - Miele, Lucio

PY - 2015/12/1

Y1 - 2015/12/1

N2 - p53 and Notch-1 play important roles in breast cancer biology. Notch-1 inhibits p53 activity in cervical and breast cancer cells. Conversely, p53 inhibits Notch activity in T-cells but stimulates it in human keratinocytes. Notch co-activator MAML1 binds p53 and functions as a p53 co-activator. We studied the regulation of Notch signaling by p53 in MCF-7 cells and normal human mammary epithelial cells (HMEC). Results show that overexpression of p53 or activation of endogenous p53 with Nutlin-3 inhibits Notch-dependent transcriptional activity and Notch target expression in a dose-dependent manner. This effect could be partially rescued by transfection of MAML1 but not p300. Standard and quantitative co-immunoprecipitation experiments readily detected a complex containing p53 and Notch-1 in MCF-7 cells. Formation of this complex was inhibited by dominant negative MAML1 (DN-MAML1) and stimulated by wild-type MAML1. Standard and quantitative far-Western experiments showed a complex including p53, Notch-1, and MAML1. Chromatin immunoprecipitation (ChIP) experiments showed that p53 can associate with Notch-dependent HEY1 promoter and this association is inhibited by DN-MAML1 and stimulated by wild-type MAML1. Our data support a model in which p53 associates with the Notch transcriptional complex (NTC) in a MAML1-dependent fashion, most likely through a p53-MAML1 interaction. In our cellular models, the effect of this association is to inhibit Notch-dependent transcription. Our data suggest that p53-null breast cancers may lack this Notch-modulatory mechanism, and that therapeutic strategies that activate wild-type p53 can indirectly cause inhibition of Notch transcriptional activity.

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