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Type: Journal article
Title: Promoter I of the ovine acetyl-CoA carboxylase-a gene: an E-box motif at - 114 in the proximal promoter binds upstream stimulatory factor (USF)-1 and USF-2 and acts as an insulin-response sequence in differentiating adipocytes
Author: Travers, M.
Vallance, A.
Gourlay, H.
Gill, C.
Klein, I.
Bottema, C.
Barber, M.
Citation: Biochemical Journal, 2001; 359(Part 2):273-284
Publisher: Portland Press
Issue Date: 2001
ISSN: 0264-6021
Statement of
Maureen T. Travers, Amanda J. Vallance, Helen T. Gourlay, Clare A. Gill, Izabella Klein, Cynthia B. K. Bottema and Michael C. Barber
Abstract: Acetyl-CoA carboxylase-a (ACC-a) plays a central role in co-ordinating de novo fatty acid synthesis in animal tissues. We have characterized the regulatory region of the ovine ACC-a gene. Three promoters, PI, PII and PIII, are dispersed throughout 50kb of genomic DNA. Expression from PI is limited to adipose tissue and liver. Sequence comparison of the proximal promoters of ovine and mouse PIs demonstrates high nucleotide identity and that they are characterized by a TATA box at -29, C/EBP (CCAAT enhancer-binding protein)-binding motifs and multiple E-box motifs. A 4.3kb ovine PI-luciferase reporter construct is insulin-responsive when transfected into differentiated ovine adipocytes, whereas when this construct is transfected into ovine preadipocytes and HepG2 cells the construct is inactive and is not inducible by insulin. By contrast, transfection of a construct corresponding to 132bp of the proximal promoter linked to a luciferase reporter is active and inducible by insulin in all three cell systems. Insulin signalling to the -132bp construct in differentiated ovine adipocytes involves, in part, an E-box motif at -114. Upstream stimulatory factor (USF)-1 and USF-2, but not sterol regulatory element-binding protein 1 (SREBP-1), are major components of protein complexes that bind this E-box motif. Activation of the 4.3kb PI construct in differentiated ovine adipocytes is associated with endogenous expression of PI transcripts throughout differentiation; PI transcripts are not detectable by RNase-protection assay in ovine preadipocytes, HepG2 cells or 3T3-F442A adipocytes. These data indicate the presence of repressor motifs in PI that are required to be de-repressed during adipocyte differentiation to allow induction of the promoter by insulin.
Keywords: Adipose tissue
HepG2 cells
insulin signalling
DOI: 10.1042/bj3590273
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