ISOLATION AND CHARACTERIZATION OF THE ANDROGEN-DEPENDENT MOUSE CYSTEINE-RICH SECRETORY PROTEIN-3 (CRISP-3) GENE

The mRNA for cysteine-rich secretory protein-3 (CRISP-3) was originall y identified in the mouse salivary gland as an androgen-dependent tran script, and is closely related to CRISP-1 and CRISP-2 which are abunda ntly expressed in the epididymis and testis respectively. Overlapping phage clones encompassing the entire length of the CRISP-3 gene were i solated from a lambda EMBL3 genomic library and analysed. DNA sequenci ng revealed that the gene consisted of eight exons ranging between 55 and 740 bp in size, and seven introns. All exon-intron junctions confo rmed to the GT/AG rule established for eukaryotic genes. The length of the introns was determined by PCR and was found to vary between 1.0 a nd 3.7 kb, indicating that the gene spans over 20 kb of the mouse geno me. Primer extension allowed the mapping of the major transcription in itiation site to an adenine located at the appropriate position downst ream of a bona fide TATA box, in a region corresponding well to the eu karyotic consensus sequence. Over 800 bp of CRISP-3 promoter region we re determined and two regions almost exactly matching the androgen-res ponsive element consensus RGWACANNNTGTWCY detected. In addition, seque nces described in the Drosophila melanogaster Sgs-3 gene as being invo lved in its salivary gland-specific expression as well as two putative OTF- and GATA-binding elements were also found.