ISOLATION AND CHARACTERIZATION OF THE ANDROGEN-DEPENDENT MOUSE CYSTEINE-RICH SECRETORY PROTEIN-1 (CRISP-1) GENE

In mice, cysteine-rich secretory protein-1 (CRISP-1) is mainly found i n the epididymis and also, to a lesser extent, in the salivary gland o f males, where androgens control its expression. We have now isolated and characterized overlapping phage clones covering the entire length of the CRISP-1 gene. DNA sequencing revealed that the gene is organize d into eight exons, ranging between 55 and 748 bp in size, and seven i ntrons. All exon-intron junctions conformed to the GT/AG rule establis hed for eukaryotic genes. The intron length, as determined by PCR, var ied between 1.05 and 4.0 kb so that the CRISP-1 gene spans over 20 kb of the mouse genome. The transcription-initiation site was determined by primer extension and localized at the expected distance downstream of a consensus TATA box. Approximately 3.7 kb of the CRISP-1 promoter region were isolated and sequenced, and several stretches fitting the androgen-responsive element consensus were found. Those that most rese mbled the consensus were analysed by electrophoretic mobility-shift as say and found to form specific complexes with the liganded androgen re ceptor in vitro, but with different affinities. Putative binding eleme nts for the transcription factors Oct, GATA, PEA3, CF1, AP-1 and AP-3 were also found in the promoter region.