Molecular organization of the rat glia-derived nexin/protease nexin-1 promoter.
The first three exons and the promoter of rat glia-derived nexin, also called protease nexin-1 (GDN/PN-1), have been identified through analysis of rat genomic clones. A 1.6 kilobase (kb) fragment containing 105 base pairs of the first exon and 5'-flanking sequences was sequenced. The 5'-flanking sequence and the first exon were found to be GC-rich, indicating that the 5' region of the rat GDN/PN-1 gene resides within a CpG island. A TATA box-like sequence, but no CAAT box, was found. The rat GDN/PN-1 promoter contains five SP1 consensus sites, four consensus sites for the MyoD1 transcription factor, and one binding site for the transcription factors NGFI-A, NGFI-C, Krox-20, and Wilms tumor factor. The presence of these consensus sequences is consistent with the known expression pattern of GDN/PN-1. Primer extension and RNase protection assays identified one transcriptional start site. The 1.6 kb promoter fragment cloned in a reporter plasmid was found to induce firefly luciferase expression in a cell-specific manner. A positive regulatory element is localized in the region -1545 to -389. In vitro CpG methylation blocked transcription from the GDN/PN-1 promoter in rat hepatoma cells but not in C6 rat glioma cells.
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Friedrich Miescher Institute, Basel, Switzerland.
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D016564:Amyloid beta-Protein Precursor; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification; D015226:Dinucleoside Phosphates / Q000378:metabolism; D005091:Exons; D008745:Methylation; D008969:Molecular Sequence Data; D009457:Neuroglia / Q000378:metabolism*; D015849:Plasminogen Inactivators / Q000235:genetics*; D011401:Promoter Regions, Genetic; D058255:Protease Nexins; D051381:Rats; D011956:Receptors, Cell Surface; D015183:Restriction Mapping; D014158:Transcription, Genetic
D016564:Amyloid beta-Protein Precursor; D002352:Carrier Proteins; D004247:DNA; D015226:Dinucleoside Phosphates; D015849:Plasminogen Inactivators; D058255:Protease Nexins; D011956:Receptors, Cell Surface; C015772:CpG dinucleotide