TY - JOUR
T1 - C/EBP-beta drives expression of the nutritionally regulated promoter IA of the acetyl-CoA carboxylase-alpha gene in cattle
AU - Shi, Xuanming
AU - Liu, Shuzhen
AU - Metges, Cornelia C.
AU - Seyfert, Hans Martin
N1 - Funding Information:
We are grateful to Angelika Deike and Bärbel Pletz for technical assistance and Dr. Wolfgang Tomek for help with the ECL detection system. X.S. and S.L. contributed equally to this work and the results are integral part of either of their doctoral thesis. This work is supported by the Deutsche Forschungsgemeinschaft through grant Se 326/13 .
PY - 2010/8
Y1 - 2010/8
N2 - Acetyl-CoA carboxylase-alpha (ACC-α) 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-α gene expression in cattle, we established vectors expressing full length (FL) or N-terminally truncated (δN) variants of the C/EBP factors (α, -β, -γ, and -ε) in mammalian cells. Using nuclear extracts of cells expressing the δN-C/EBP factors we determined in electrophoretic mobility shift assays that C/EBPα, -β and -ε, but not C/EBPγ may directly bind to the cognate C/EBP-binding site in the immediate ACC-α PIA. Co-transfection analyses of the various FL-C/EBP expression vectors together with a reporter gene driven by the ACC-α-PIA promoter demonstrated that C/EBPβ has the strongest activation potential. Hence, activity of this factor may be a key regulator of ACC-α-expression in lipogenic tissues.
AB - Acetyl-CoA carboxylase-alpha (ACC-α) 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-α gene expression in cattle, we established vectors expressing full length (FL) or N-terminally truncated (δN) variants of the C/EBP factors (α, -β, -γ, and -ε) in mammalian cells. Using nuclear extracts of cells expressing the δN-C/EBP factors we determined in electrophoretic mobility shift assays that C/EBPα, -β and -ε, but not C/EBPγ may directly bind to the cognate C/EBP-binding site in the immediate ACC-α PIA. Co-transfection analyses of the various FL-C/EBP expression vectors together with a reporter gene driven by the ACC-α-PIA promoter demonstrated that C/EBPβ has the strongest activation potential. Hence, activity of this factor may be a key regulator of ACC-α-expression in lipogenic tissues.
KW - ACC-alpha
KW - C/EBP
KW - Cattle
KW - Gene expression
KW - Isoform
KW - Lipid synthesis
UR - http://www.scopus.com/inward/record.url?scp=77955653938&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77955653938&partnerID=8YFLogxK
U2 - 10.1016/j.bbagrm.2010.07.002
DO - 10.1016/j.bbagrm.2010.07.002
M3 - Article
C2 - 20637321
AN - SCOPUS:77955653938
SN - 1874-9399
VL - 1799
SP - 561
EP - 567
JO - Biochimica et Biophysica Acta - Gene Regulatory Mechanisms
JF - Biochimica et Biophysica Acta - Gene Regulatory Mechanisms
IS - 8
ER -