Nuclear factors AF-1 and C/EBP bind to the human ApoB gene promoter and modulate its transcriptional activity in hepatic cells.
ApoB-100, the major protein of low density lipoprotein, is produced primarily in the liver. The apoB promoter region from -86 to -52 binds two hepatic nuclear proteins. The protein that binds to the sequence between -86 to -61, called AF-1, was characterized and also shown to interact with the apoCIII, apoA-I, and apoCII promoters. We now show that the second protein, which by DNase I footprinting binds to the sequence between -69 and -52, is the heat stable hepatic factor C/EBP. The binding of this protein is competed by a known C/EBP-binding sequence in the transthyretin gene enhancer (TRR oligo 3). In addition, the protein protects the same region in DNase I footprinting as purified C/EBP. Mutants that affect AF-1 or C/EBP binding to the apoB promoter were created. These mutants were analyzed by the gel mobility shift assay and transient transfections into HepG2 cells. We have demonstrated that the two factors bind to overlapping sites and that they modulate apoB transcriptional activity.
Metzger
S
S
Laboratory of Biochemical Genetics and Metabolism, Rockefeller University, New York, New York 10021.
Leff
T
T
Breslow
J L
JL
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000235:genetics*; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism*; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014162:Transfection; D014452:Ubiquitins / Q000378:metabolism*
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D015345:Oligonucleotide Probes; D014452:Ubiquitins