Involvement of estrogen receptor variant ER-α36, not GPR30, in nongenomic estrogen signaling

Lianguo Kang, Xintian Zhang, Yan Xie, Yaping Tu, Dong Wang, Zhenming Liu, Zhao Yi Wang

Research output: Contribution to journalArticle

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Abstract

Accumulating evidence suggested that an orphan G protein-coupled receptor (GPR)30, mediates nongenomic responses to estrogen. The present study was performed to investigate the molecular mechanisms underlying GPR30 function.Wefound that knockdown of GPR30 expression in breast cancer SK-BR-3 cells down-regulated the expression levels of estrogen receptor (ER)-α36, a variant of ER-α. Introduction of a GPR30 expression vector into GPR30 nonexpressing cells induced endogenous ER-α36 expression, and cotransfection assay demonstrated that GPR30 activated the promoter activity of ER-α36 via an activator protein 1 binding site. Both 17β-estradiol (E2) and G1, a compound reported to be a selective GPR30 agonist, increased the phosphorylation levels of the MAPK/ERK1/2 in SK-BR-3 cells, which could be blocked by an anti-ER-α36-specific antibody against its ligand-binding domain. G1 induced activities mediated by ER-α36, such as transcription activation activity of a VP16-ER-α36 fusion protein and activation of the MAPK/ERK1/2 in ER-α36-expressing cells. ER-α36-expressing cells, but not the nonexpressing cells, displayed high-affinity, specific E2 and G1 binding, and E2- and G1-induced intracellular Ca2+ mobilization only in ER-α36 expressing cells. Taken together, our results demonstrated that previously reported activities of GPR30 in response to estrogen were through its ability to induce ER-α36 expression. The selective G protein-coupled receptor (GPR)30 agonist G1 actually interacts with ER-α36. Thus, the ER-α variant ER-α36, not GPR30, is involved in nongenomic estrogen signaling.

Original languageEnglish
Pages (from-to)709-721
Number of pages13
JournalMolecular Endocrinology
Volume24
Issue number4
DOIs
StatePublished - Apr 2010

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Estrogen Receptors
Estrogens
G-Protein-Coupled Receptors
Transcription Factor AP-1
Protein Binding
Transcriptional Activation
Estradiol
Binding Sites
Phosphorylation
Breast Neoplasms
Ligands

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Endocrinology

Cite this

Involvement of estrogen receptor variant ER-α36, not GPR30, in nongenomic estrogen signaling. / Kang, Lianguo; Zhang, Xintian; Xie, Yan; Tu, Yaping; Wang, Dong; Liu, Zhenming; Wang, Zhao Yi.

In: Molecular Endocrinology, Vol. 24, No. 4, 04.2010, p. 709-721.

Research output: Contribution to journalArticle

Kang, Lianguo ; Zhang, Xintian ; Xie, Yan ; Tu, Yaping ; Wang, Dong ; Liu, Zhenming ; Wang, Zhao Yi. / Involvement of estrogen receptor variant ER-α36, not GPR30, in nongenomic estrogen signaling. In: Molecular Endocrinology. 2010 ; Vol. 24, No. 4. pp. 709-721.
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abstract = "Accumulating evidence suggested that an orphan G protein-coupled receptor (GPR)30, mediates nongenomic responses to estrogen. The present study was performed to investigate the molecular mechanisms underlying GPR30 function.Wefound that knockdown of GPR30 expression in breast cancer SK-BR-3 cells down-regulated the expression levels of estrogen receptor (ER)-α36, a variant of ER-α. Introduction of a GPR30 expression vector into GPR30 nonexpressing cells induced endogenous ER-α36 expression, and cotransfection assay demonstrated that GPR30 activated the promoter activity of ER-α36 via an activator protein 1 binding site. Both 17β-estradiol (E2) and G1, a compound reported to be a selective GPR30 agonist, increased the phosphorylation levels of the MAPK/ERK1/2 in SK-BR-3 cells, which could be blocked by an anti-ER-α36-specific antibody against its ligand-binding domain. G1 induced activities mediated by ER-α36, such as transcription activation activity of a VP16-ER-α36 fusion protein and activation of the MAPK/ERK1/2 in ER-α36-expressing cells. ER-α36-expressing cells, but not the nonexpressing cells, displayed high-affinity, specific E2 and G1 binding, and E2- and G1-induced intracellular Ca2+ mobilization only in ER-α36 expressing cells. Taken together, our results demonstrated that previously reported activities of GPR30 in response to estrogen were through its ability to induce ER-α36 expression. The selective G protein-coupled receptor (GPR)30 agonist G1 actually interacts with ER-α36. Thus, the ER-α variant ER-α36, not GPR30, is involved in nongenomic estrogen signaling.",
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