Contribution of a nuclear factor 1 binding site to the glucocorticoid regulation of the cytosolic aspartate aminotransferase gene promoter.
Abstract
Two regions of the cAspAT gene promoter mediate the glucocorticoid regulation of this gene in the Fao hepatoma cell line. The proximal region was localized by deletion studies and stable transfections in the Fao cells to the sequence -553/-398. This region includes the glucocorticoid-responsive element (GRE) A sequence, which consists of two overlapping GREs and which can mediate the glucocorticoid regulation of a heterologous promoter. DNase I footprinting studies have shown that a site 80 base pairs upstream of the GRE A was protected by liver and brain nuclear extracts (site P8). The binding was displaced by an excess of an oligonucleotide containing a typical NF1 binding site and by NF1-specific antibodies. In electrophoretic mobility shift assay using the P8 oligonucleotide as a probe, several complexes were formed. Most complexes were common to liver and brain but were less abundant when testis extracts were used. At least one complex was specific to the liver. All complexes, with the exception of two, were competed for by the NF1 oligonucleotide. Furthermore, the sequence of the P8 site showed a 7/9-base pair homology with a typical NF1 site. A mutation of the P8 site, which prevents the binding of NF1-like proteins to i...Continue Reading
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