Activation of SIRT1 by resveratrol represses transcription of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) by deacetylating hepatic nuclear factor 4α

Jianqi Yang, Xiaoying Kong, Maria Emilia S Martins-Santos, Gabriela Aleman, Ernestine Chaco, George E. Liu, Shwu Yuan Wu, David Samols, Parvin Hakimi, Cheng Ming Chiang, Richard W. Hanson

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Abstract

The SIRT1 activators isonicotinamide (IsoNAM), resveratrol, fisetin, and butein repressed transcription of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (PEPCK-C). An evolutionarily conserved binding site for hepatic nuclear factor (HNF) 4α (-272/ -252) was identified, which was required for transcriptional repression of the PEPCK-C gene promoter caused by these compounds. This site contains an overlapping AP-1 binding site and is adjacent to the C/EBP binding element (-248/-234); the latter is necessary for hepatic transcription of PEPCK-C. AP-1 competed with HNF4α for binding to this site and also decreased HNF4α stimulation of transcription from the PEPCK-C gene promoter. Chromatin immunoprecipitation experiments demonstrated that HNF4α and AP-1, but not C/EBPβ, reciprocally bound to this site prior to and after treating HepG2 cells with IsoNAM. IsoNAM treatment resulted in deacetylation of HNF4α, which decreased its binding affinity to the PEPCK-C gene promoter. In HNF4α-null Chinese hamster ovary cells, IsoNAM and resveratrol failed to repress transcription from the PEPCK-C gene promoter; overexpression of HNF4α in Chinese hamster ovary cells re-established transcriptional inhibition. Exogenous SIRT1 expression repressed transcription, whereas knock-down of SIRT1 by RNA interference reversed this effect. Iso-NAM decreased the level of mRNA for PEPCK-C but had no effect on mRNA for glucose-6-phosphatase in AML12 mouse hepatocytes. We conclude that SIRT1 activation inhibited transcription of the gene for PEPCK-C in part by deacetylation of HNF4α. However, SIRT1 deacetylation of other key regulatory proteins that control PEPCK-C gene transcription also likely contributed to the inhibitory effect.

Original languageEnglish (US)
Pages (from-to)27042-27053
Number of pages12
JournalJournal of Biological Chemistry
Volume284
Issue number40
DOIs
StatePublished - Oct 2 2009

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Phosphoenolpyruvate Carboxykinase (GTP)
Transcription
Genes
Chemical activation
Liver
Transcription Factor AP-1
Binding Sites
Cricetulus
Ovary
Cells
Glucose-6-Phosphatase
Messenger RNA
Chromatin Immunoprecipitation
Hep G2 Cells
RNA Interference
resveratrol
Transcriptional Activation
Chromatin
Hepatocytes
RNA

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Activation of SIRT1 by resveratrol represses transcription of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) by deacetylating hepatic nuclear factor 4α. / Yang, Jianqi; Kong, Xiaoying; Martins-Santos, Maria Emilia S; Aleman, Gabriela; Chaco, Ernestine; Liu, George E.; Wu, Shwu Yuan; Samols, David; Hakimi, Parvin; Chiang, Cheng Ming; Hanson, Richard W.

In: Journal of Biological Chemistry, Vol. 284, No. 40, 02.10.2009, p. 27042-27053.

Research output: Contribution to journalArticle

Yang, Jianqi ; Kong, Xiaoying ; Martins-Santos, Maria Emilia S ; Aleman, Gabriela ; Chaco, Ernestine ; Liu, George E. ; Wu, Shwu Yuan ; Samols, David ; Hakimi, Parvin ; Chiang, Cheng Ming ; Hanson, Richard W. / Activation of SIRT1 by resveratrol represses transcription of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) by deacetylating hepatic nuclear factor 4α. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 40. pp. 27042-27053.
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abstract = "The SIRT1 activators isonicotinamide (IsoNAM), resveratrol, fisetin, and butein repressed transcription of the gene for the cytosolic form of phosphoenolpyruvate carboxykinase (GTP) (PEPCK-C). An evolutionarily conserved binding site for hepatic nuclear factor (HNF) 4α (-272/ -252) was identified, which was required for transcriptional repression of the PEPCK-C gene promoter caused by these compounds. This site contains an overlapping AP-1 binding site and is adjacent to the C/EBP binding element (-248/-234); the latter is necessary for hepatic transcription of PEPCK-C. AP-1 competed with HNF4α for binding to this site and also decreased HNF4α stimulation of transcription from the PEPCK-C gene promoter. Chromatin immunoprecipitation experiments demonstrated that HNF4α and AP-1, but not C/EBPβ, reciprocally bound to this site prior to and after treating HepG2 cells with IsoNAM. IsoNAM treatment resulted in deacetylation of HNF4α, which decreased its binding affinity to the PEPCK-C gene promoter. In HNF4α-null Chinese hamster ovary cells, IsoNAM and resveratrol failed to repress transcription from the PEPCK-C gene promoter; overexpression of HNF4α in Chinese hamster ovary cells re-established transcriptional inhibition. Exogenous SIRT1 expression repressed transcription, whereas knock-down of SIRT1 by RNA interference reversed this effect. Iso-NAM decreased the level of mRNA for PEPCK-C but had no effect on mRNA for glucose-6-phosphatase in AML12 mouse hepatocytes. We conclude that SIRT1 activation inhibited transcription of the gene for PEPCK-C in part by deacetylation of HNF4α. However, SIRT1 deacetylation of other key regulatory proteins that control PEPCK-C gene transcription also likely contributed to the inhibitory effect.",
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AU - Kong, Xiaoying

AU - Martins-Santos, Maria Emilia S

AU - Aleman, Gabriela

AU - Chaco, Ernestine

AU - Liu, George E.

AU - Wu, Shwu Yuan

AU - Samols, David

AU - Hakimi, Parvin

AU - Chiang, Cheng Ming

AU - Hanson, Richard W.

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