The 5'-flanking region of the alpha 2MR/LRP gene contains an enhancer-like cluster of Sp1 binding sites.
We have sequenced a genomic DNA fragment containing the promoter and 5'-flanking region of the alpha 2MR/LRP. A cluster of five Sp1 sites situated over 600 base pairs away from the putative transcription start site doubles the activity of the promoter. A similar increase in activity was observed when this region was replaced by the SV40 enhancer, but the presence of both the cluster of Sp1 sites and SV40 enhancer gave no more transcription than either region alone. Within the previously described promoter region we have shown that only the most proximal Sp1 binding site influences transcription in CHO cells. The Sp 1 site situated 346 bp upstream of the putative transcription start site and previously described DNAse protection footprints had no effect on promoter activity in CHO cells. We also describe an NRF-1 binding site situated 143 bp upstream of the putative transcription start site. Deletion of the central 4 bp of this site caused a 60% decrease in transcription. No sterol regulatory (SRE-1) sites, used in the LDL receptor promoter for control of expression by cholesterol, were found in the alpha 2MR/LRP 5'-flanking region. However, one SRE-1 site was identified in the 5'-untranslated region of alpha 2MR/LRP.
Gaëta
B A
BA
Heart Research Institute, Camperdown, Sydney NSW, Australia.
Borthwick
I
I
Stanley
K K
KK
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011971:Receptors, Immunologic / Q000235:genetics*; D016415:Sequence Alignment; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D016329:Sp1 Transcription Factor