title: The histone demethylase PHF2 promotes fat cell differentiation as an epigenetic activator of both C/EBPα and C/EBPδ.
abstract: Histone modifications on major transcription factor target genes are one of the major regulatory mechanisms controlling adipogenesis. Plant homeodomain finger 2 (PHF2) is a Jumonji domain-containing protein and is known to demethylate the histone H3K9, a repressive gene marker. To better understand the function of PHF2 in adipocyte differentiation, we constructed stable PHF2 knock-down cells by using the mouse pre-adipocyte cell line 3T3-L1. When induced with adipogenic media, PHF2 knock-down cells showed reduced lipid accumulation compared to control cells. Differential expression using a cDNA microarray revealed significant reduction of metabolic pathway genes in the PHF2 knock-down cell line after differentiation. The reduced expression of major transcription factors and adipokines was confirmed with reverse transcription- quantitative polymerase chain reaction and Western blotting. We further performed co-immunoprecipitation analysis of PHF2 with four major adipogenic transcription factors, and we found that CCATT/enhancer binding protein (C/EBP)α and C/EBPδ physically interact with PHF2. In addition, PHF2 binding to target gene promoters was confirmed with a chromatin immunoprecipitation experiment. Finally, histone H3K9 methylation markers on the PHF2-binding sequences were increased in PHF2 knock-down cells after differentiation. Together, these results demonstrate that PHF2 histone demethylase controls adipogenic gene expression during differentiation.
authors.affiliation: Ischemic/Hypoxic Disease Institute, Seoul National University College of Medicine, Seoul 110-799, Korea.
authors.forename: Kyoung-Hwa
authors.initials: KH
authors.lastname: Lee
authors.affiliation: Departments of Biomedical Sciences, Seoul National University College of Medicine, Seoul 110-799, Korea.
authors.forename: Uk-Il
authors.initials: UI
authors.lastname: Ju
authors.affiliation: Departments of Biomedical Sciences, Seoul National University College of Medicine, Seoul 110-799, Korea.
authors.forename: Jung-Yup
authors.initials: JY
authors.lastname: Song
authors.affiliation: Ischemic/Hypoxic Disease Institute, Seoul National University College of Medicine, Seoul 110-799, Korea ; Departments of Biomedical Sciences, Seoul National University College of Medicine, Seoul 110-799, Korea ; Departments of Physiology, Seoul National University College of Medicine, Seoul 110-799, Korea.
authors.forename: Yang-Sook
authors.initials: YS
authors.lastname: Chun
mesh_terms: D016475:3T3 Cells; D017667:Adipocytes / Q000502:physiology*; D050156:Adipogenesis / Q000235:genetics*; D000818:Animals; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics*; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics*; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D055785:Gene Knockdown Techniques; D056466:Histone Demethylases / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D050356:Lipid Metabolism / Q000235:genetics; D051379:Mice; D046228:Microarray Analysis; D013234:Stem Cells / Q000502:physiology*
chemical_list: D022782:CCAAT-Enhancer-Binding Protein-beta; D022762:CCAAT-Enhancer-Binding Proteins; C530718:CEBPA protein, mouse; C545013:Cebpb protein, mouse; D006657:Histones; D018398:Homeodomain Proteins; D056466:Histone Demethylases; C000598540:PHF2 protein, mouse
keywords: PHF2; adipogenesis; epigenetic regulation; histone demethylase; transcriptional control