Human nuclear pregnane X receptor cross-talk with CREB to repress cAMP activation of the glucose-6-phosphatase gene

Susumu Kodama, Rick Moore, Yukio Yamamoto, Masahiko Negishi

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

The nuclear PXR (pregnane X receptor) was originally characterized as a key transcription factor that activated hepatic genes encoding drug-metabolizing enzymes. We have now demonstrated that PXR also represses glucagon-activated transcription of the G6Pase (glucose-6-phosphatase) gene by directly binding to CREB [CRE (cAMP-response element)-binding protein]. Adenoviral-mediated expression of human PXR (hPXR) and its activation by rifampicin strongly repressed cAMP-dependent induction of the endogenous G6Pase gene in Huh7 cells. Using the -259 bp G6Pase promoter construct in cell-based transcription assays, repression by hPXR of PKA (cAMP-dependent protein kinase)-mediated promoter activation was delineated to CRE sites. GST (glutathione transferase) pull-down and immunoprecipitation assays were employed to show that PXR binds directly to CREB, while gel-shift assays were used to demonstrate that this binding prevents CREB interaction with the CRE. These results are consistent with the hypothesis that PXR represses the transcription of the G6Pase gene by inhibiting the DNA-binding ability of CREB. In support of this hypothesis, treatment with the mouse PXR activator PCN (pregnenolone 16α-carbonitrile) repressed cAMP-dependent induction of the G6Pase gene in primary hepatocytes prepared from wild-type, but not from PXR-knockout, mice, and also in the liver of fasting wild-type, but not PXR-knockout, mice. Moreover, ChIP (chromatin immunoprecipitation) assays were performed to show a decreased CREB binding to the G6Pase promoter in fasting wild-type mice after PCN treatment. Thus drug activation of PXR can repress the transcriptional activity of CREB, down-regulating gluconeogenesis.

Original languageEnglish
Pages (from-to)373-381
Number of pages9
JournalBiochemical Journal
Volume407
Issue number3
DOIs
Publication statusPublished - Nov 1 2007
Externally publishedYes

Fingerprint

Receptor Cross-Talk
Cyclic AMP Response Element-Binding Protein
Glucose-6-Phosphatase
Genes
Chemical activation
Assays
Transcription
Pregnenolone Carbonitrile
Knockout Mice
Fasting
pregnane X receptor
Gene encoding
Aptitude
Gluconeogenesis
Chromatin Immunoprecipitation
Liver
Rifampin
Cyclic AMP-Dependent Protein Kinases
Glutathione Transferase
Glucagon

Keywords

  • cAMP-response element-binding protein (CREB)
  • Glucagon
  • Gluconeogenesis
  • Glucose-6-phosphatase (G6Pase)
  • Insulin
  • Pregnane X receptor (PXR)

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Human nuclear pregnane X receptor cross-talk with CREB to repress cAMP activation of the glucose-6-phosphatase gene. / Kodama, Susumu; Moore, Rick; Yamamoto, Yukio; Negishi, Masahiko.

In: Biochemical Journal, Vol. 407, No. 3, 01.11.2007, p. 373-381.

Research output: Contribution to journalArticle

Kodama, Susumu ; Moore, Rick ; Yamamoto, Yukio ; Negishi, Masahiko. / Human nuclear pregnane X receptor cross-talk with CREB to repress cAMP activation of the glucose-6-phosphatase gene. In: Biochemical Journal. 2007 ; Vol. 407, No. 3. pp. 373-381.
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