The transcriptional repressor GATAD2B mediates progesterone receptor suppression of myometrial contractile gene expression

Chien Cheng Chen, Alina P. Montalbano, Imran Hussain, Wan Ru Lee, Carole R. Mendelson

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The mechanisms whereby progesterone (P4), acting via the progesterone receptor (PR), inhibits proinflammatory/contractile gene expression during pregnancy are incompletely defined. Using immortalized human myometrial (hTERT-HM) cells stably expressing wild-type PR-A or PR-B (PRWT), we found that P4 significantly inhibited IL-1 induction of the NF-B target genes, COX-2 and IL-8. P4–PRWT transrepression occurred at the level of transcription initiation and was mediated by decreased recruitment of NF-B p65 and RNA polymerase II to COX-2 and IL-8 promoters. However, in cells stably expressing a PR-A or PR-B DNA-binding domain mutant (PRmDBD), P4-mediated transrepression was significantly reduced, suggesting a critical role of the PR DBD. ChIP analysis of hTERT-HM cells stably expressing PRWT or PRmDBD revealed that P4 treatment caused equivalent recruitment of PRWT and PRmDBD to COX-2 and IL-8 promoters, suggesting that PR inhibitory effects were not mediated by its direct DNA binding. Using immunoprecipitation, followed by MS, we identified a transcriptional repressor, GATA zinc finger domain–containing 2B (GATAD2B), that interacted strongly with PRWT but poorly with PRmDBD. P4 treatment of PRWT hTERT-HM cells caused enhanced recruitment of endogenous GATAD2B to COX-2 and IL-8 promoters. Further, siRNA knockdown of endogenous GATAD2B significantly reduced P4–PRWT transrepression of COX-2 and IL-8. Notably, GATAD2B expression was significantly decreased in pregnant mouse and human myometrium during labor. Our findings suggest that GATAD2B serves as an important mediator of P4–PR suppression of proinflammatory and contractile genes during pregnancy. Decreased GATAD2B expression near term may contribute to the decline in PR function, leading to labor.

Original languageEnglish (US)
Pages (from-to)12560-12576
Number of pages17
JournalJournal of Biological Chemistry
Volume292
Issue number30
DOIs
StatePublished - 2017

Fingerprint

Zinc Fingers
Progesterone Receptors
Gene expression
Zinc
Interleukin-8
Gene Expression
Genes
Personnel
Pregnancy
RNA Polymerase II
Myometrium
DNA
Transcription
Interleukin-1
Immunoprecipitation
Small Interfering RNA
Progesterone

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The transcriptional repressor GATAD2B mediates progesterone receptor suppression of myometrial contractile gene expression. / Chen, Chien Cheng; Montalbano, Alina P.; Hussain, Imran; Lee, Wan Ru; Mendelson, Carole R.

In: Journal of Biological Chemistry, Vol. 292, No. 30, 2017, p. 12560-12576.

Research output: Contribution to journalArticle

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abstract = "The mechanisms whereby progesterone (P4), acting via the progesterone receptor (PR), inhibits proinflammatory/contractile gene expression during pregnancy are incompletely defined. Using immortalized human myometrial (hTERT-HM) cells stably expressing wild-type PR-A or PR-B (PRWT), we found that P4 significantly inhibited IL-1 induction of the NF-B target genes, COX-2 and IL-8. P4–PRWT transrepression occurred at the level of transcription initiation and was mediated by decreased recruitment of NF-B p65 and RNA polymerase II to COX-2 and IL-8 promoters. However, in cells stably expressing a PR-A or PR-B DNA-binding domain mutant (PRmDBD), P4-mediated transrepression was significantly reduced, suggesting a critical role of the PR DBD. ChIP analysis of hTERT-HM cells stably expressing PRWT or PRmDBD revealed that P4 treatment caused equivalent recruitment of PRWT and PRmDBD to COX-2 and IL-8 promoters, suggesting that PR inhibitory effects were not mediated by its direct DNA binding. Using immunoprecipitation, followed by MS, we identified a transcriptional repressor, GATA zinc finger domain–containing 2B (GATAD2B), that interacted strongly with PRWT but poorly with PRmDBD. P4 treatment of PRWT hTERT-HM cells caused enhanced recruitment of endogenous GATAD2B to COX-2 and IL-8 promoters. Further, siRNA knockdown of endogenous GATAD2B significantly reduced P4–PRWT transrepression of COX-2 and IL-8. Notably, GATAD2B expression was significantly decreased in pregnant mouse and human myometrium during labor. Our findings suggest that GATAD2B serves as an important mediator of P4–PR suppression of proinflammatory and contractile genes during pregnancy. Decreased GATAD2B expression near term may contribute to the decline in PR function, leading to labor.",
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AB - The mechanisms whereby progesterone (P4), acting via the progesterone receptor (PR), inhibits proinflammatory/contractile gene expression during pregnancy are incompletely defined. Using immortalized human myometrial (hTERT-HM) cells stably expressing wild-type PR-A or PR-B (PRWT), we found that P4 significantly inhibited IL-1 induction of the NF-B target genes, COX-2 and IL-8. P4–PRWT transrepression occurred at the level of transcription initiation and was mediated by decreased recruitment of NF-B p65 and RNA polymerase II to COX-2 and IL-8 promoters. However, in cells stably expressing a PR-A or PR-B DNA-binding domain mutant (PRmDBD), P4-mediated transrepression was significantly reduced, suggesting a critical role of the PR DBD. ChIP analysis of hTERT-HM cells stably expressing PRWT or PRmDBD revealed that P4 treatment caused equivalent recruitment of PRWT and PRmDBD to COX-2 and IL-8 promoters, suggesting that PR inhibitory effects were not mediated by its direct DNA binding. Using immunoprecipitation, followed by MS, we identified a transcriptional repressor, GATA zinc finger domain–containing 2B (GATAD2B), that interacted strongly with PRWT but poorly with PRmDBD. P4 treatment of PRWT hTERT-HM cells caused enhanced recruitment of endogenous GATAD2B to COX-2 and IL-8 promoters. Further, siRNA knockdown of endogenous GATAD2B significantly reduced P4–PRWT transrepression of COX-2 and IL-8. Notably, GATAD2B expression was significantly decreased in pregnant mouse and human myometrium during labor. Our findings suggest that GATAD2B serves as an important mediator of P4–PR suppression of proinflammatory and contractile genes during pregnancy. Decreased GATAD2B expression near term may contribute to the decline in PR function, leading to labor.

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