Structure, organization and regulation of a rat cysteine proteinase inhibitor-encoding gene

During postnatal development, submandibular glands of rats produce the secretory protein, cystatin S (CysS), which belongs to family 2 of the mammalian cysteine proteinase inhibitor superfamily. While the rat CysS gene is not expressed in the salivary glands of adult rats, it can be induced by isoproterenol (IPR), which acts via beta-adrenergic receptor/adenylate cyclase/cyclic AMP (cAMP) mechanisms. In addition, IPR-induction of CysS mRNA in submandibular glands is more pronounced in females than in males, at both prepuberal and mature ages. These results suggest that sex hormones may participate in the regulation of the rat CysS gene via estrogen-responsive elements (ERE), and IPR induction of this gene supports the hypothesis that cAMP-responsive elements (CRE) may also play a role in regulating CysS gene expression. We have isolated, sequenced and characterized the complete gene. The CysS gene contains three exons interrupted by two intervening sequences, with consensus splice junctions. The transcription start point is 73 nucleotides upstream from the start codon which is surrounded by a typical Kozak sequence. CCAAT and TATA boxes are present in the 5'-flanking region of the CysS gene. This region also contains several possible regulatory elements that resemble those of other eukaryotic genes, i.e., ERE, CRE, and glucocorticoid-responsive elements. The first intron sequence contains other potential CRE highly homologous to those found in the IPR-inducible mouse and hamster proline-rich-protein-encoding genes.