title: Fatmass and obesity associated (FTO) gene regulates gluconeogenesis in  chicken embryo fibroblast cells.
abstract: Fat mass and obesity-associated (FTO) gene was found to be associated with energy homeostasis in mammals, yet the function of chicken FTO is less clear. In this study, chicken embryo fibroblast cells (DF-1) were transiently transfected to over-express (FTO(+)) or to knockdown (FTO−) the chicken FTO gene and were used for the functional analysis. FTO expression was significantly augmented in FTO(+) cells while depressed in FTO(−) cells (P < 0.05). FTO(+) cells had significantly lower glucose yet higher lactic acid (LD) concentrations (P < 0.05) in the culture media, which was associated with significantly up-regulated (P < 0.05) mRNA expression of the rate-limiting gluconeogenic enzymes, glucose-6-phosphatase (G6PC) and the phosphoenolpyruvate carboxykinase-mitochondrial (PEPCK-m). The protein content and enzyme activity of G6PC were also significantly higher (P < 0.05) in FTO(+) cells. Moreover, CCAAT/enhancer-binding protein-beta (C/EBP-beta) and cAMP responsive element binding protein 1 (CREB1), which were found to transcriptionally regulate the expression of G6PC, were increased at the level of both mRNA (P < 0.05) and protein (P < 0.05) in FTO(+)cells. ChIP analysis revealed significantly higher (P < 0.05) binding of C/EBP-beta and phospho-CREB1 to G6PC gene promoter in FTO(+) cells. In addition, the interaction of FTO and C/EBP-beta was significantly enhanced (P < 0.05) in FTO+ cells. Opposite changes in G6PC expression and regulation were observed in FTO(−) cells. Our results indicate that chicken FTO regulates gluconeogenesis in DF-1 cells through enhanced transcriptional regulation of G6PC gene by C/EBP-beta and phospho-CREB1.
authors.forename: Feng
authors.initials: F
authors.lastname: Guo
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Chunxiao
authors.initials: C
authors.lastname: Zhang
authors.forename: Song
authors.initials: S
authors.lastname: Wang
authors.forename: Yingdong
authors.initials: Y
authors.lastname: Ni
authors.forename: Ruqian
authors.initials: R
authors.lastname: Zhao
mesh_terms: D000818:Animals; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival; D002642:Chick Embryo; D003470:Culture Media; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D005943:Gluconeogenesis / Q000235:genetics*; D005947:Glucose / Q000378:metabolism; D005952:Glucose-6-Phosphatase / Q000235:genetics / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D022782:CCAAT-Enhancer-Binding Protein-beta; D003470:Culture Media; D017362:Cyclic AMP Response Element-Binding Protein; D012333:RNA, Messenger; D019344:Lactic Acid; D006003:Glycogen; D006899:Mixed Function Oxygenases; D005952:Glucose-6-Phosphatase; D005947:Glucose