C/EBP-beta drives expression of the nutritionally regulated promoter IA of the acetyl-CoA carboxylase-alpha gene in cattle.
Acetyl-CoA carboxylase-alpha (ACC-alpha) is the rate-limiting enzyme for de novo fatty acid synthesis. Among the four promoters expressing the bovine gene, promoter IA (PIA) is dominantly active and nutritionally regulated in lipogenic tissues. CCAAT/enhancer binding proteins are crucially involved in regulating the activity of this promoter. We examine here, which member of this family of transcription factors is most important for promoter activation. To differentiate the individual contribution of different members of the C/EBP family transcription factors controlling the ACC-alpha gene expression in cattle, we established vectors expressing full length (FL) or N-terminally truncated (DeltaN) variants of the C/EBP factors (alpha, -beta, -delta, and -epsilon) in mammalian cells. Using nuclear extracts of cells expressing the DeltaN-C/EBP factors we determined in electrophoretic mobility shift assays that C/EBPalpha, -beta and -epsilon, but not C/EBPdelta may directly bind to the cognate C/EBP-binding site in the immediate ACC-alpha PIA. Co-transfection analyses of the various FL-C/EBP expression vectors together with a reporter gene driven by the ACC-alpha-PIA promoter demonstrated that C/EBPbeta has the strongest activation potential. Hence, activity of this factor may be a key regulator of ACC-alpha-expression in lipogenic tissues.
Shi
Xuanming
X
Research Units for Molecular Biology, Leibniz Institute for Farm Animal Biology (FBN), Wilhelm-Stahl-Allee 2, D-18196 Dummerstorf, Germany.
Liu
Shuzhen
S
Metges
Cornelia C
CC
Seyfert
Hans-Martin
HM
D000103:Acetyl-CoA Carboxylase / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D008321:Mammary Glands, Animal / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
D022782:CCAAT-Enhancer-Binding Protein-beta; D012333:RNA, Messenger; D000103:Acetyl-CoA Carboxylase