title: Hepatocyte nuclear factor (HNF)-4alpha-driven epigenetic silencing of the human PED gene.
abstract: OBJECTIVE
  Overexpression of PED (also known as PEA15) determines insulin resistance and impaired insulin secretion and may contribute to progression toward type 2 diabetes. Recently, we found that the transcription factor hepatocyte nuclear factor (HNF)-4alpha binds to PED promoter and represses its transcription. However, the molecular details responsible for regulation of PED gene remain unclear.
  
  
  METHODS
  Here we used gain and loss of function approaches to investigate the hypothesis that HNF-4alpha controls chromatin remodelling at the PED promoter in human cell lines.
  
  
  RESULTS
  HNF-4alpha production and binding induce chromatin remodelling at the -250 to 50 region of PED, indicating that remodelling is limited to two nucleosomes located at the proximal promoter. Chromatin immunoprecipitation assays also revealed concomitant HNF-4alpha-induced deacetylation of histone H3 at Lys9 and Lys14, and increased dimethylation of histone H3 at Lys9. The latter was followed by reduction of histone H3 Lys4 dimethylation. HNF-4alpha was also shown to target the histone deacetylase complex associated with silencing mediator of retinoic acid and thyroid hormone receptor, both at the PED promoter, and at GRB14 and USP21 regulatory regions, leading to a reduction of mRNA levels. Moreover, HNF-4alpha silencing and PED overexpression were accompanied by a significant reduction of hepatic glycogen content.
  
  
  CONCLUSIONS
  These results show that HNF-4alpha serves as a scaffold protein for histone deacetylase activities, thereby inhibiting liver expression of genes including PED. Dysregulation of these mechanisms may lead to upregulation of the PED gene in type 2 diabetes.
authors.affiliation: Dipartimento di Biologia e Patologia Cellulare e Molecolare L. Califano & Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università di Napoli Federico II, Via Sergio Pansini, 5, Naples, 80131, Italy. pungaro@ieos.cnr.it
authors.forename: P
authors.initials: P
authors.lastname: Ungaro
authors.forename: R
authors.initials: R
authors.lastname: Teperino
authors.forename: P
authors.initials: P
authors.lastname: Mirra
authors.forename: M
authors.initials: M
authors.lastname: Longo
authors.forename: M
authors.initials: M
authors.lastname: Ciccarelli
authors.forename: G A
authors.initials: GA
authors.lastname: Raciti
authors.forename: C
authors.initials: C
authors.lastname: Nigro
authors.forename: C
authors.initials: C
authors.lastname: Miele
authors.forename: P
authors.initials: P
authors.lastname: Formisano
authors.forename: F
authors.initials: F
authors.lastname: Beguinot
mesh_terms: D000107:Acetylation; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D015153:Blotting, Western; D042002:Chromatin Assembly and Disassembly / Q000235:genetics / Q000502:physiology; D047369:Chromatin Immunoprecipitation; D044127:Epigenesis, Genetic / Q000235:genetics / Q000502:physiology*; D056945:Hep G2 Cells; D051557:Hepatocyte Nuclear Factor 4 / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009707:Nucleosomes / Q000235:genetics; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D051017:Apoptosis Regulatory Proteins; D051557:Hepatocyte Nuclear Factor 4; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D009707:Nucleosomes; C092039:PEA15 protein, human; D010750:Phosphoproteins
references.citation: Cell. 2007 Feb 23;128(4):693-705
references.pmid: 17320507
references.citation: J Neurochem. 1995 Mar;64(3):1016-25
references.pmid: 7861130
references.citation: Diabetes. 2000 Mar;49(3):424-30
references.pmid: 10868964
references.citation: Cytokine. 1999 Apr;11(4):305-12
references.pmid: 10328870
references.citation: J Biol Chem. 2007 Mar 23;282(12):8648-57
references.pmid: 17227770
references.citation: Mol Cell Biol. 2004 Jun;24(11):5005-15
references.pmid: 15143191
references.citation: Genes Dev. 2000 May 1;14(9):1048-57
references.pmid: 10809664
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):20021-6
references.pmid: 19066220
references.citation: Diabetologia. 2006 Dec;49(12):3058-66
references.pmid: 17021921
references.citation: EMBO J. 1998 Jul 15;17(14):3858-66
references.pmid: 9670003
references.citation: Genes Dev. 1990 Dec;4(12B):2353-65
references.pmid: 2279702
references.citation: Mol Cell Biol. 2007 Sep;27(17):5933-48
references.pmid: 17591692
references.citation: J Clin Invest. 2008 Jun;118(6):2316-24
references.pmid: 18464933
references.citation: Cell. 2002 Feb 22;108(4):465-74
references.pmid: 11909518
references.citation: J Cell Sci. 2003 Jun 1;116(Pt 11):2117-24
references.pmid: 12730288
references.citation: Biochem Biophys Res Commun. 1997 Dec 29;241(3):664-9
references.pmid: 9434765
references.citation: Cell. 2004 Jun 11;117(6):721-33
references.pmid: 15186774
references.citation: Mol Cell Biol. 2002 Mar;22(6):1626-38
references.pmid: 11865043
references.citation: Genes Dev. 2006 Jun 1;20(11):1405-28
references.pmid: 16751179
references.citation: Diabetologia. 1999 Feb;42(2):139-45
references.pmid: 10064092
references.citation: Cell Mol Life Sci. 2000 Sep;57(10):1388-98
references.pmid: 11078018
references.citation: J Clin Invest. 2005 Apr;115(4):1006-15
references.pmid: 15761495
references.citation: Diabetes. 2001 Jun;50(6):1244-52
references.pmid: 11375323
references.citation: Metabolism. 2008 Oct;57 Suppl 2:S27-31
references.pmid: 18803962
references.citation: Science. 2004 Feb 27;303(5662):1378-81
references.pmid: 14988562
references.citation: Nat Rev Genet. 2007 Apr;8(4):253-62
references.pmid: 17363974
references.citation: Nutr Metab Cardiovasc Dis. 2005 Apr;15(2):109-17
references.pmid: 15871859
references.citation: Nature. 1998 Feb 19;391(6669):815-8
references.pmid: 9486655
references.citation: Science. 2007 Jun 1;316(5829):1331-6
references.pmid: 17463246
references.citation: Genes Dev. 2000 Feb 15;14(4):464-74
references.pmid: 10691738
references.citation: Mol Endocrinol. 2006 Jan;20(1):1-13
references.pmid: 16002433
references.citation: J Pediatr Endocrinol Metab. 2001 Sep-Oct;14(8):1085-91
references.pmid: 11592564
references.citation: Nat Genet. 2003 Jul;34(3):292-6
references.pmid: 12808453
references.citation: Nat Genet. 2003 Mar;33 Suppl:245-54
references.pmid: 12610534
references.citation: PLoS Genet. 2006 Sep 22;2(9):e158
references.pmid: 17002501
references.citation: Diabetologia. 2004 Aug;47(8):1354-9
references.pmid: 15258738
references.citation: J Immunol. 2001 Oct 15;167(8):4494-503
references.pmid: 11591776
references.citation: Proc Natl Acad Sci U S A. 2006 Oct 24;103(43):15782-7
references.pmid: 17043231
references.citation: Nat Rev Mol Cell Biol. 2006 Jun;7(6):437-47
references.pmid: 16723979
references.citation: J Biol Chem. 1998 Nov 20;273(47):30847-30850
references.pmid: 9812974
references.citation: Nature. 2000 Jan 6;403(6765):41-5
references.pmid: 10638745
references.citation: Mol Cell Biol. 2001 Feb;21(4):1393-403
references.pmid: 11158324
references.citation: Curr Opin Cell Biol. 2003 Apr;15(2):172-83
references.pmid: 12648673
references.citation: Nature. 2007 Feb 22;445(7130):881-5
references.pmid: 17293876
references.citation: J Biol Chem. 2008 Nov 7;283(45):30970-9
references.pmid: 18765665
references.citation: Diabetes. 2008 Apr;57(4):1048-56
references.pmid: 18162503
references.citation: Genes Dev. 2004 Jun 1;18(11):1263-71
references.pmid: 15175259
references.citation: Clin Genet. 2008 Feb;73(2):185-7
references.pmid: 18028455
references.citation: Trends Genet. 2008 Dec;24(12):613-21
references.pmid: 18952314
references.citation: Nature. 2002 Sep 26;419(6905):407-11
references.pmid: 12353038
references.citation: Diabetes. 2007 Dec;56(12):3112-7
references.pmid: 17827402
references.citation: Diabetes Care. 2008 Jul;31(7):1422-6
references.pmid: 18375420