Abstract
The ability of the human insulin receptor promoter to direct expression of a linked chloramphenicol acetyltransferase gene was assessed in transient transfections into HepG2 and Hela cells. A 5'-deletional analysis of the promoter showed that regions between -646 and -489 were important for the activity of the proximal promoter. In addition, a possible negative regulatory element was identified between -1311 and -877 and a positive element between -1823 and -1311. DNase I footprint and gel retardation analysis showed that multiple factors bind to the human insulin receptor promoter. In particular, DNase I protection patterns were observed over the Sp1 sites at -620 to -599 and -438 to -392, a TC box at -533, four homopyrimidine/homopurine sites clustered around -1150, and a site at -1420 that contains the motif TGGCCC which has been shown to bind the liver-specific transcription factor LF-A1.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Base Sequence
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Binding Sites
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Carcinoma, Hepatocellular
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Cell Line
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Chloramphenicol O-Acetyltransferase / biosynthesis
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Chloramphenicol O-Acetyltransferase / genetics
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Chromosome Deletion
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Cloning, Molecular
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DNA-Binding Proteins / metabolism
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Deoxyribonuclease I
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Gene Expression Regulation*
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HeLa Cells
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Hepatocyte Nuclear Factor 1
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Hepatocyte Nuclear Factor 1-alpha
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Hepatocyte Nuclear Factor 1-beta
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Humans
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Liver Neoplasms
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Molecular Sequence Data
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Nuclear Proteins*
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Plasmids
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Promoter Regions, Genetic*
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Receptor, Insulin / biosynthesis
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Receptor, Insulin / genetics*
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Recombinant Fusion Proteins / biosynthesis
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Restriction Mapping
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Transcription Factors / metabolism*
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Transcription, Genetic*
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Transfection
Substances
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DNA-Binding Proteins
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HNF1A protein, human
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HNF1B protein, human
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Hepatocyte Nuclear Factor 1-alpha
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Nuclear Proteins
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Recombinant Fusion Proteins
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Transcription Factors
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Hepatocyte Nuclear Factor 1
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Hepatocyte Nuclear Factor 1-beta
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Chloramphenicol O-Acetyltransferase
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Receptor, Insulin
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Deoxyribonuclease I
Associated data
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GENBANK/J05043
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GENBANK/M84424
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GENBANK/M92919
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GENBANK/M92920
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GENBANK/M93702
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GENBANK/M93703
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GENBANK/M93704
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GENBANK/M93705
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GENBANK/M94144
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GENBANK/M94316