Propionate induces the bovine cytosolic phosphoenolpyruvate carboxykinase promoter activity.
Cytosolic phosphoenolpyruvate carboxykinase (PCK1) is a critical enzyme within the metabolic networks for gluconeogenesis, hepatic energy metabolism, and tricarboxylic acid cycle function, and is controlled by several transcription factors including hepatic nuclear factor 4α (HNF4α). The primary objective of the present study was to determine whether propionate regulates bovine PCK1 transcription. The second objective was to determine the action of cyclic AMP (cAMP), glucocorticoids, and insulin, hormonal cues known to modulate glucose metabolism, on bovine PCK1 transcriptional activity. The proximal promoter of the bovine PCK1 gene was ligated to a Firefly luciferase reporter and transfected into H4IIE hepatoma cells. Cells were exposed to treatments for 23 h and luciferase activity was determined in cell lysates. Activity of the PCK1 promoter was linearly induced by propionate, and maximally increased 7-fold with 2.5 mM propionate, which was not muted by 100 nM insulin. Activity of the PCK1 promoter was increased 1-fold by either 1.0 mM cAMP or 5.0µM dexamethasone, and 2.2-fold by their combination. Induction by cAMP and dexamethasone was repressed 50% by 100 nM insulin. Propionate, cAMP, and dexamethasone acted synergistically to induce the PCK1 promoter activity. Propionate-responsive regions, identified by 5' deletion analysis, were located between -1,238 and -409 bp and between -85 and +221 bp. Deletions of the core sequences of the 2 putative HNF4α sites decreased the responsiveness to propionate by approximately 40%. These data indicate that propionate regulates its own metabolism through transcriptional stimulation of the bovine PCK1 gene. This induction is mediated, in part, by the 2 putative HNF4α binding sites in the bovine PCK1 promoter.
Department of Animal Sciences, Purdue University, West Lafayette, IN 47907.
Qian
Q
Zhang
Department of Animal Sciences, Purdue University, West Lafayette, IN 47907.
Stephanie L
SL
Koser
Department of Animal Sciences, Purdue University, West Lafayette, IN 47907. Electronic address: sdonkin@purdue.edu.
Shawn S
SS
Donkin
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D010728:Phosphoenolpyruvate; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011422:Propionates; D014158:Transcription, Genetic
D011422:Propionates; D010728:Phosphoenolpyruvate; D010729:Phosphoenolpyruvate Carboxykinase (GTP)
PCK1; hormone; promoter; short-chain fatty acid