title: Transcription factor assisted loading and enhancer dynamics dictate the hepatic fasting response.
abstract: Fasting elicits transcriptional programs in hepatocytes leading to glucose and ketone production. This transcriptional program is regulated by many transcription factors (TFs). To understand how this complex network regulates the metabolic response to fasting, we aimed at isolating the enhancers and TFs dictating it. Measuring chromatin accessibility revealed that fasting massively reorganizes liver chromatin, exposing numerous fasting-induced enhancers. By utilizing computational methods in combination with dissecting enhancer features and TF cistromes, we implicated four key TFs regulating the fasting response: glucocorticoid receptor (GR), cAMP responsive element binding protein 1 (CREB1), peroxisome proliferator activated receptor alpha (PPARA), and CCAAT/enhancer binding protein beta (CEBPB). These TFs regulate fuel production by two distinctly operating modules, each controlling a separate metabolic pathway. The gluconeogenic module operates through assisted loading, whereby GR doubles the number of sites occupied by CREB1 as well as enhances CREB1 binding intensity and increases accessibility of CREB1 binding sites. Importantly, this GR-assisted CREB1 binding was enhancer-selective and did not affect all CREB1-bound enhancers. Single-molecule tracking revealed that GR increases the number and DNA residence time of a portion of chromatin-bound CREB1 molecules. These events collectively result in rapid synergistic gene expression and higher hepatic glucose production. Conversely, the ketogenic module operates via a GR-induced TF cascade, whereby PPARA levels are increased following GR activation, facilitating gradual enhancer maturation next to PPARA target genes and delayed ketogenic gene expression. Our findings reveal a complex network of enhancers and TFs that dynamically cooperate to restore homeostasis upon fasting.
authors.affiliation: Laboratory of Receptor Biology and Gene Expression, The National Cancer Institute, The National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Ido
authors.initials: I
authors.lastname: Goldstein
authors.affiliation: Laboratory of Receptor Biology and Gene Expression, The National Cancer Institute, The National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Songjoon
authors.initials: S
authors.lastname: Baek
authors.affiliation: Laboratory of Receptor Biology and Gene Expression, The National Cancer Institute, The National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Diego M
authors.initials: DM
authors.lastname: Presman
authors.affiliation: Laboratory of Receptor Biology and Gene Expression, The National Cancer Institute, The National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Ville
authors.initials: V
authors.lastname: Paakinaho
authors.affiliation: Laboratory of Receptor Biology and Gene Expression, The National Cancer Institute, The National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Erin E
authors.initials: EE
authors.lastname: Swinstead
authors.affiliation: Laboratory of Receptor Biology and Gene Expression, The National Cancer Institute, The National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Gordon L
authors.initials: GL
authors.lastname: Hager
mesh_terms: D000818:Animals; D001665:Binding Sites; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics / Q000378:metabolism*; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism*; D004742:Enhancer Elements, Genetic; D005215:Fasting / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism*; D007659:Ketones / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D047493:PPAR alpha / Q000235:genetics / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation
chemical_list: D022782:CCAAT-Enhancer-Binding Protein-beta; C545013:Cebpb protein, mouse; D002843:Chromatin; C492004:Creb1 protein, mouse; D017362:Cyclic AMP Response Element-Binding Protein; D007659:Ketones; D047493:PPAR alpha; D011965:Receptors, Glucocorticoid; D005947:Glucose