Localization of a polyunsaturated fatty acid response region in stearoyl-CoA desaturase gene 1.
Polyunsaturated fatty acids (PUFA) repress stearoyl-CoA desaturase gene 1 (SCD1) expression in liver and adipose tissues. We used HepG2 cells to localize genetic regulatory elements for PUFA in the SCD1 5'-flanking region. A chimeric reporter gene construct containing the 4.3 kb SCD1 putative promoter was transiently transfected into HepG2 cells, which were then treated with various fatty acids. We observed greater than 60% repression of transcription with 18:3n - 3 and 75% repression with 20:4n - 6 and 20:5n - 3. No significant change was seen with 18:0. Using smaller SCD1 chimeric constructs, we localized the genetic regulatory region to a 237 bp sequence within the SCD1 proximal promoter. DNA mobility shift analysis with HepG2 and mouse liver nuclear extracts demonstrated specific binding of nuclear proteins to this region. Mobility shift analysis with nuclear extract from 3T3-L1 adipocytes showed a similar pattern of protein binding. Competitive DNA mobility shift analysis identified a 60 bp region containing sites that specifically bind and compete for nuclear proteins. This region conferred responsiveness to PUFA when placed in a heterologous promoter. A homologous region in the stearoyl-CoA desaturase gene 2 (SCD2) promoter also mediated PUFA-specific repression in transfection experiments. These data suggest that a common transcriptional mechanism may exist in liver and adipose tissues for inhibition of lipogenesis by PUFA.
Waters
K M
KM
Department of Biochemistry, The University of Wisconsin-Madison, 53706, USA.
Miller
C W
CW
Ntambi
J M
JM
D017667:Adipocytes / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D006801:Humans; D008099:Liver / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000494:pharmacology; D013230:Stearoyl-CoA Desaturase / Q000235:genetics*; D015533:Transcriptional Activation
D005231:Fatty Acids, Unsaturated; D009687:Nuclear Proteins; D012097:Repressor Proteins; D013230:Stearoyl-CoA Desaturase