Hepatocyte nuclear factor (HNF)-4alpha-driven epigenetic silencing of the human PED gene.
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.
Ungaro
P
P
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
Teperino
R
R
Mirra
P
P
Longo
M
M
Ciccarelli
M
M
Raciti
G A
GA
Nigro
C
C
Miele
C
C
Formisano
P
P
Beguinot
F
F
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
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
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