Progesterone receptor-nfκb complex formation is required for progesterone-induced nfκb nuclear translocation and binding onto the p53 promoter

Sung Po Hsu, Ho Ching Yang, Chun Ting Kuo, Heng Ching Wen, Li Ching Chen, Yen Nien Huo, Wen Sen Lee

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We previously demonstrated that progesterone (P4) up-regulates p53 expression in human umbilical venous endothelial cells (HUVECs) through P4 receptor (PR) activation of extranuclear signaling pathways. However, the involvement of nuclear PR in P4-increased p53 expression is still unclear. Here, the molecular mechanism underlying PR-regulated p53 expression in HUVECs was investigated. Treatment with P4 increased nuclear factor of κ light polypeptide gene enhancer inB-cells inhibitor, α phosphorylation (IκBα and nuclear factor-κB (NFκB) nuclear translocation. Interestingly, P4 also increased PR-A, but not PR-B, nuclear translocation in HUVECs. Immunoprecipitation assay illustrated that P4 increased the formation of PR-A-NFκB complex in both the cytosol and the nucleus of HUVEC. Chromatin immunoprecipitation assay showed an interaction between PR and the NFκB binding motif on the p53 promoter. Ablation of the NFκB binding motif in the p53 promoter completely abolished P4-increased p53 promoter activity. In the absence of P4, overexpression of NFκB did not increase NFκB nuclear translocation. In contrast, treatment of NFκB-overexpressing HUVECs with P4 for only 4 hours, which is much shorter than the time (21.5 h) required for P4-induced IκBα phosphorylation, increased NFκB nuclear translocation. Blockade ofPR activity abolished this effect. Taken together, these results uncover a novel role of PR for P4-induced NFκB nuclear translocation and suggest that PR-A-NFκB complex formation is required for NFκB nuclear translocation and binding onto the p53 promoter in HUVECs. Our data indicate that both nuclear and extranuclear signaling pathways of PR are involved in P4-regulated p53 expression in HUVECs.

Original languageEnglish
Pages (from-to)291-300
Number of pages10
JournalEndocrinology
Volume156
Issue number1
DOIs
Publication statusPublished - Jan 1 2015

Fingerprint

Umbilicus
Progesterone Receptors
Progesterone
Endothelial Cells
Cytoplasmic and Nuclear Receptors
Phosphorylation
Chromatin Immunoprecipitation
Immunoprecipitation
Cytosol
Up-Regulation
Light
Peptides

ASJC Scopus subject areas

  • Endocrinology
  • Medicine(all)

Cite this

Progesterone receptor-nfκb complex formation is required for progesterone-induced nfκb nuclear translocation and binding onto the p53 promoter. / Hsu, Sung Po; Yang, Ho Ching; Kuo, Chun Ting; Wen, Heng Ching; Chen, Li Ching; Huo, Yen Nien; Lee, Wen Sen.

In: Endocrinology, Vol. 156, No. 1, 01.01.2015, p. 291-300.

Research output: Contribution to journalArticle

@article{ff3a52ea275249a7b96fe09c4b6cbd73,
title = "Progesterone receptor-nfκb complex formation is required for progesterone-induced nfκb nuclear translocation and binding onto the p53 promoter",
abstract = "We previously demonstrated that progesterone (P4) up-regulates p53 expression in human umbilical venous endothelial cells (HUVECs) through P4 receptor (PR) activation of extranuclear signaling pathways. However, the involvement of nuclear PR in P4-increased p53 expression is still unclear. Here, the molecular mechanism underlying PR-regulated p53 expression in HUVECs was investigated. Treatment with P4 increased nuclear factor of κ light polypeptide gene enhancer inB-cells inhibitor, α phosphorylation (IκBα and nuclear factor-κB (NFκB) nuclear translocation. Interestingly, P4 also increased PR-A, but not PR-B, nuclear translocation in HUVECs. Immunoprecipitation assay illustrated that P4 increased the formation of PR-A-NFκB complex in both the cytosol and the nucleus of HUVEC. Chromatin immunoprecipitation assay showed an interaction between PR and the NFκB binding motif on the p53 promoter. Ablation of the NFκB binding motif in the p53 promoter completely abolished P4-increased p53 promoter activity. In the absence of P4, overexpression of NFκB did not increase NFκB nuclear translocation. In contrast, treatment of NFκB-overexpressing HUVECs with P4 for only 4 hours, which is much shorter than the time (21.5 h) required for P4-induced IκBα phosphorylation, increased NFκB nuclear translocation. Blockade ofPR activity abolished this effect. Taken together, these results uncover a novel role of PR for P4-induced NFκB nuclear translocation and suggest that PR-A-NFκB complex formation is required for NFκB nuclear translocation and binding onto the p53 promoter in HUVECs. Our data indicate that both nuclear and extranuclear signaling pathways of PR are involved in P4-regulated p53 expression in HUVECs.",
author = "Hsu, {Sung Po} and Yang, {Ho Ching} and Kuo, {Chun Ting} and Wen, {Heng Ching} and Chen, {Li Ching} and Huo, {Yen Nien} and Lee, {Wen Sen}",
year = "2015",
month = "1",
day = "1",
doi = "10.1210/en.2014-1629",
language = "English",
volume = "156",
pages = "291--300",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "1",

}

TY - JOUR

T1 - Progesterone receptor-nfκb complex formation is required for progesterone-induced nfκb nuclear translocation and binding onto the p53 promoter

AU - Hsu, Sung Po

AU - Yang, Ho Ching

AU - Kuo, Chun Ting

AU - Wen, Heng Ching

AU - Chen, Li Ching

AU - Huo, Yen Nien

AU - Lee, Wen Sen

PY - 2015/1/1

Y1 - 2015/1/1

N2 - We previously demonstrated that progesterone (P4) up-regulates p53 expression in human umbilical venous endothelial cells (HUVECs) through P4 receptor (PR) activation of extranuclear signaling pathways. However, the involvement of nuclear PR in P4-increased p53 expression is still unclear. Here, the molecular mechanism underlying PR-regulated p53 expression in HUVECs was investigated. Treatment with P4 increased nuclear factor of κ light polypeptide gene enhancer inB-cells inhibitor, α phosphorylation (IκBα and nuclear factor-κB (NFκB) nuclear translocation. Interestingly, P4 also increased PR-A, but not PR-B, nuclear translocation in HUVECs. Immunoprecipitation assay illustrated that P4 increased the formation of PR-A-NFκB complex in both the cytosol and the nucleus of HUVEC. Chromatin immunoprecipitation assay showed an interaction between PR and the NFκB binding motif on the p53 promoter. Ablation of the NFκB binding motif in the p53 promoter completely abolished P4-increased p53 promoter activity. In the absence of P4, overexpression of NFκB did not increase NFκB nuclear translocation. In contrast, treatment of NFκB-overexpressing HUVECs with P4 for only 4 hours, which is much shorter than the time (21.5 h) required for P4-induced IκBα phosphorylation, increased NFκB nuclear translocation. Blockade ofPR activity abolished this effect. Taken together, these results uncover a novel role of PR for P4-induced NFκB nuclear translocation and suggest that PR-A-NFκB complex formation is required for NFκB nuclear translocation and binding onto the p53 promoter in HUVECs. Our data indicate that both nuclear and extranuclear signaling pathways of PR are involved in P4-regulated p53 expression in HUVECs.

AB - We previously demonstrated that progesterone (P4) up-regulates p53 expression in human umbilical venous endothelial cells (HUVECs) through P4 receptor (PR) activation of extranuclear signaling pathways. However, the involvement of nuclear PR in P4-increased p53 expression is still unclear. Here, the molecular mechanism underlying PR-regulated p53 expression in HUVECs was investigated. Treatment with P4 increased nuclear factor of κ light polypeptide gene enhancer inB-cells inhibitor, α phosphorylation (IκBα and nuclear factor-κB (NFκB) nuclear translocation. Interestingly, P4 also increased PR-A, but not PR-B, nuclear translocation in HUVECs. Immunoprecipitation assay illustrated that P4 increased the formation of PR-A-NFκB complex in both the cytosol and the nucleus of HUVEC. Chromatin immunoprecipitation assay showed an interaction between PR and the NFκB binding motif on the p53 promoter. Ablation of the NFκB binding motif in the p53 promoter completely abolished P4-increased p53 promoter activity. In the absence of P4, overexpression of NFκB did not increase NFκB nuclear translocation. In contrast, treatment of NFκB-overexpressing HUVECs with P4 for only 4 hours, which is much shorter than the time (21.5 h) required for P4-induced IκBα phosphorylation, increased NFκB nuclear translocation. Blockade ofPR activity abolished this effect. Taken together, these results uncover a novel role of PR for P4-induced NFκB nuclear translocation and suggest that PR-A-NFκB complex formation is required for NFκB nuclear translocation and binding onto the p53 promoter in HUVECs. Our data indicate that both nuclear and extranuclear signaling pathways of PR are involved in P4-regulated p53 expression in HUVECs.

UR - http://www.scopus.com/inward/record.url?scp=84919779773&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84919779773&partnerID=8YFLogxK

U2 - 10.1210/en.2014-1629

DO - 10.1210/en.2014-1629

M3 - Article

C2 - 25353185

AN - SCOPUS:84919779773

VL - 156

SP - 291

EP - 300

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 1

ER -