Isolation and characterization of the androgen-dependent mouse cysteine-rich secretory protein-1 (CRISP-1) gene

Abstract

In mice, cysteine-rich secretory protein-1 (CRISP-1) is mainly found in the epididymis and also, to a lesser extent, in the salivary gland of 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 organized into eight exons, ranging between 55 and 748 bp in size, and seven introns. All exon-intron junctions conformed to the GT/AG rule established for eukaryotic genes. The intron length, as determined by PCR, varied 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 resembled the consensus were analysed by electrophoretic mobility-shift assay and found to form specific complexes with the liganded androgen receptor in vitro, but with different affinities. Putative binding elements for the transcription factors Oct, GATA, PEA3, CF1. AP-1 and AP-3 were also found in the promoter region.

Involvement of Androgen Receptor in 17β-Estradiol-Induced Cell Proliferation in Rat Uterus1

“…10928-042) in a Gene Amp PCR system 2400 (Perkin-Elmer, Norwalk, CT). Primers, together with their annealing temperatures and the number of cycles, were IGF-1: forward, 5Ј-CACATCTCTTCTACCTGGCA-3Ј, reverse, 5Ј-TGAGTCTTGGGCATGTCAGT (56ЊC, 35 cycles) [24]; Crisp: forward, 5Ј-TGTTCCTGGCTGCTGTATTG-3Ј, reverse, 5Ј-AAGACCACGATGCAG-GGTAA-3Ј (58ЊC, 35 cycles); ␤-actin: forward, 5Ј-GGGCACAGTGTGGGT-GAC-3Ј, reverse, 5Ј-CTGGCACCACACCTTCTAC-3Ј (56ЊC, 15 cycles). PCR products were resolved in a 1.5% agarose gel and stained with ethidium bromide, and DNA bands from triplicate reactions were quantified using Fluor-S MultiImager (Bio-Rad).…”

Structure, alternative splicing and chromosomal localization of the cystatin-related epididymal spermatogenic gene

“…cystatins, including the glutathione peroxidase-like [26] and CRISP-1 genes [27]. These elements have the potential to adopt alternative DNA conformations that might direct the binding of transcription factors or other regulatory proteins to sites in the promoter [28].…”

Expression and secretion of rat SPAM1(2B1 or PH-20) in the epididymis: role of testicular lumicrine factors

“…Since epididymally-expressed genes are androgenregulated (Robaire and Viger, 1995;Schwidetzky et al, 1997) and expression of SPAM1 in humans and mice is accompanied by the presence of putative AREs in the promoter (Zhang and Martin-DeLeon, 2001;Evans et al, 2003), we searched the database for such elements in the rat promoter region. The presence of a putative site with 100% homology to the consensus at position y229 to y225, among other potential sites, is consistent with the finding of epididymal expression of the mRNA for SPAM1 in the rat.…”