Journal of Biological Chemistry · 1998 · 388 citations · 41 references
Life SciencesCell BiologyMolecular BiologyGene ExpressionGene RegulationTranscriptional RegulationGene Expression RegulationGlycan StructureGlycosylation PathwaysHormonal Gene RegulationSteroid Hormone SignalingTranslational RegulationGene Regulatory Circuit MotifsGlucocorticoid ReceptorsGlucocorticoid SignalingGlycoprotein StructureGlycosaminoglycan BiosynthesisRegulatory Dna MotifsRegulatory MotifsTrabecular Meshwork Physiology
Expression of the trabecular meshwork inducible glucocorticoid response (TIGR) gene progressively increases from barely detectable levels to greater than 2% of total cellular mRNA over 10 days exposure of trabecular meshwork (TM) cells to dexamethasone. Cycloheximide blocked most of the TIGR mRNA induction, suggesting a requirement for ongoing protein synthesis. The genomic structure of TIGR (approximately 20 kilobases) consists of 3 exons, and a 5-kilobase promoter region that contains 13 predicted hormone response elements, including several glucocorticoid regulatory elements, and other potentially important regulatory motifs. TIGR cDNA encodes an olfactomedin-related glycoprotein of 504 amino acids with motifs for N- and O-linked glycosylation, glycosaminoglycan initiation, hyaluronic acid binding, and leucine zippers. Recombinant TIGR (rTIGR) showed oligomerization and specific binding to TM cells. Anti-rTIGR antibody detected multiple translational/post-translational forms of TIGR produced by the cells (including secreted 66 kDa/55 kDa glycoproteins/proteins in the media and 55 kDa cellular proteins), whereas Northern blot showed a single mRNA species. The findings suggest potential mechanisms by which TIGR could obstruct the aqueous humor fluid flow and participate in the pathogenesis of glaucoma.
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A simple and very efficient method for generating cDNA libraries
Ueli Gubler, Beth J. Hoffman · Gene · 1983 · 4.6K citations