Two nuclear proteins bind to the major positive element of the apolipoprotein B gene promoter.
The promoter of the apolipoprotein B-encoding gene (apoB) contains a number of regulatory elements, which together produce a high level of expression that is restricted to two tissues: liver and intestine. In this paper we have used the gel retardation and methylation interference assays to identify two nuclear proteins, LIT1 and LIT2, which bind to the major positive element (MPE) of the apoB promoter. LIT1 is large protein, estimated to be approx. 200 kDa by gel filtration, which binds to the apoB promoter between positions of -79 and -65 bp in relation to the transcription start point. Its binding site is identical to the region responsible for cell-specific transcriptional activation. However, whereas the MPE has no influence on expression from a heterologous promoter in the non-apoB-expressing HeLa cells, these cells still contain a DNA-binding activity indistinguishable from LIT1. LIT2 binds to the apoB promoter immediately downstream from the LIT1 site. It is present in nuclear extracts from the apoB-expressing cell lines of hepatic (HepG2) and intestinal (CaCo-2) origin, but absent from HeLa cells. CCAAT/enhancer binding protein (C/EBP), expressed in bacteria, binds to the LIT2 site and produces a methylation interference pattern indistinguishable from that of LIT2. That C/EBP binds to and activates the apoB promoter in vivo, is shown by the increased chloramphenicol acetyltransferase activity observed when HepG2 cells, transfected with apoB-promoter-cat constructs, are cotransfected with a plasmid expressing c/ebp; an effect that depends on the presence in the apoB promoter of the LIT2 site.(ABSTRACT TRUNCATED AT 250 WORDS)
Carlsson
P
P
Department of Medical Biochemistry, University of Göteborg, Sweden.
Eriksson
P
P
Bjursell
G
G
D001055:Apolipoproteins B / Q000378:metabolism*; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000032:analysis*; D005786:Gene Expression Regulation; D005809:Genes, Regulator; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D014158:Transcription, Genetic; D014162:Transfection
D001055:Apolipoproteins B; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins