Interaction of CCAAT/enhancer-binding protein α and β with the rat caeruloplasmin gene promoter
| Autor: | J D Gitlin, Colin D. Bingle, R E Fleming |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Chloramphenicol O-Acetyltransferase
Transcriptional Activation Recombinant Fusion Proteins Molecular Sequence Data Gene Expression Biology Transfection Biochemistry Transcription (biology) Animals Deoxyribonuclease I Promoter Regions Genetic Molecular Biology Gene Cell Nucleus Reporter gene Binding Sites Base Sequence Ccaat-enhancer-binding proteins Oligonucleotide Ceruloplasmin Nuclear Proteins Promoter DNA Cell Biology Molecular biology Footprinting Rats DNA-Binding Proteins Liver CCAAT-Enhancer-Binding Proteins Transcription Factors Research Article |
| Zdroj: | Biochemical Journal. 294:473-479 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj2940473 |
| Popis: | To determine the mechanisms of expression of the rat caeruloplasmin gene, the promoter region was analysed by DNAase I footprinting. Using nuclear extract from rat liver, a prominent site of protein-DNA interaction was detected from -93 to -48 upstream of the caeruloplasmin gene transcription start and sequence analysis of this region revealed three potential CCAAT/enhancer-binding protein (C/EBP) consensus elements. Mobility-shift analysis using an oligonucleotide encoding this region identified specific binding of proteins from rat liver nuclear extract, and some of these complexes were supershifted using antisera to the C/EBP alpha and beta family members. Mobility-shift studies using a polypeptide encoding the DNA-binding domain of C/EBP alpha also revealed a specific interaction with this region of the caeruloplasmin promoter, and DNAase I footprinting using this polypeptide protected the identical region from -93 to -48. Co-transfection of expression plasmids encoding C/EBP alpha or a related leucine-zipper factor D-binding protein (DBP) revealed a C/EBP-specific increase in reporter gene activity in HepG2 cells transfected with caeruloplasmin-chloramphenicol acetyltransferase containing the -93 to -48 region. A similar result was obtained when these constructs were co-transfected into mouse L cells which were shown not to express the endogenous caeruloplasmin gene. Taken together, these data indicate a role for C/EBP alpha and beta in mediating transcription from the caeruloplasmin gene promoter and suggest that this region of the promoter is not responsible for tissue-specific expression. |
| Databáze: | OpenAIRE |
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