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Sterol regulatory element binding protein binds to a cis element in the promoter of the farnesyl diphosphate synthase gene
- Source :
- Proceedings of the National Academy of Sciences of the United States of America. 93(2)
- Publication Year :
- 1996
-
Abstract
- Sterol-regulated transcription of the gene for rat farnesyl diphosphate (FPP) synthase (geranyl-diphosphate:isopentenyl-diphosphate geranyltranstransferase, EC 2.5.1.10) is dependent in part on the binding of the ubiquitous transcription factor NF-Y to a 6-bp element within the proximal promoter. Current studies identify a second element in this promoter that is also required for sterol-regulated transcription in vivo. Mutation of three nucleotides (CAC) within this element blocks the 8-fold induction of FPP synthase promoter-reporter genes that normally occurs when the transfected cells are incubated in medium deprived of sterols. Gel mobility-shift assays demonstrate that the transcriptionally active 68-kDa fragment of the sterol regulatory element (SRE-1)-binding protein assays (SREBP-1) binds to an oligonucleotide containing the wild-type sequence but not to an oligonucleotide in which the CAC has been mutated. DNase 1 protection pattern (footprint) analysis indicates that SREBP-1 binds to nucleotides that include the CAC. Both the in vivo and in vitro assays are affected by mutagenesis of nucleotides adjacent to the CAC. Coexpression of SREBP with a wild-type FPP synthase promoter-reporter gene in CV-1 cells results in very high levels of reporter activity that is sterol-independent. In contrast, the reporter activity remained low when the promoter contained a mutation in the CAC trinucleotide. We conclude that sterol-regulated transcription of FPP synthase is controlled in part by the interaction of SREBP with a binding site that we have termed SRE-3. Identification of this element may prove useful in the identification of other genes that are both regulated by SREBP and involved in lipid biosynthesis.
- Subjects :
- Molecular Sequence Data
DNA Footprinting
Transfection
DNA-binding protein
Gene Expression Regulation, Enzymologic
Transcription (biology)
Genes, Reporter
Transferases
Lipid biosynthesis
Animals
Binding site
Promoter Regions, Genetic
Transcription factor
Cells, Cultured
Multidisciplinary
Alkyl and Aryl Transferases
Binding Sites
ATP synthase
biology
Base Sequence
Nuclear Proteins
Geranyltranstransferase
Molecular biology
Sterol regulatory element-binding protein
Rats
DNA-Binding Proteins
Biochemistry
Mutagenesis
biology.protein
CCAAT-Enhancer-Binding Proteins
Sterol Regulatory Element Binding Protein 1
lipids (amino acids, peptides, and proteins)
Protein Binding
Transcription Factors
Research Article
Subjects
Details
- ISSN :
- 00278424
- Volume :
- 93
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Accession number :
- edsair.doi.dedup.....575fef471e41e8311bdcc17fd7056b0d