Promoter I of the ovine acetyl-CoA carboxylase alpha 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

M. Travers, A.J. Vallance, H.T. Gourlay, C.A. Gill, I. Klein, C.B. Bottema, M.C. Barber

Research output: Contribution to journalArticle

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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.
LanguageEnglish
Pages273-284
Number of pages12
JournalBiochemical Journal
Volume359
Issue number2
Publication statusPublished - 2001

Fingerprint

Upstream Stimulatory Factors
Acetyl-CoA Carboxylase
Adipocytes
E-Box Elements
Sheep
Genes
Insulin
Luciferases
Tissue
Sterol Regulatory Element Binding Protein 1
CCAAT-Enhancer-Binding Proteins
Hep G2 Cells
TATA Box
Nucleic Acid Regulatory Sequences
Ribonucleases
Liver
Assays
Animals
Fatty Acids
Nucleotides

Keywords

  • adipose tissue
  • preadipocyte
  • HepG2 cells
  • lipogenesis
  • insulin signalling

Cite this

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title = "Promoter I of the ovine acetyl-CoA carboxylase alpha 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",
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.",
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Promoter I of the ovine acetyl-CoA carboxylase alpha 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. / Travers, M.; Vallance, A.J.; Gourlay, H.T.; Gill, C.A.; Klein, I.; Bottema, C.B.; Barber, M.C.

In: Biochemical Journal, Vol. 359, No. 2, 2001, p. 273-284.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Promoter I of the ovine acetyl-CoA carboxylase alpha 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

AU - Travers, M.

AU - Vallance, A.J.

AU - Gourlay, H.T.

AU - Gill, C.A.

AU - Klein, I.

AU - Bottema, C.B.

AU - Barber, M.C.

PY - 2001

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KW - HepG2 cells

KW - lipogenesis

KW - insulin signalling

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