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 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.
Schwidetzky
U
U
Research Laboratories of Schering AG, Berlin, Germany.
Schleuning
W D
WD
Haendler
B
B
D000728:Androgens / Q000502:physiology*; D000818:Animals; D010582:Bacteriophage lambda; D001483:Base Sequence; D003001:Cloning, Molecular; D003545:Cysteine; D004822:Epididymis / Q000737:chemistry*; D005796:Genes; D008297:Male; D008562:Membrane Glycoproteins; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011944:Receptors, Androgen / Q000737:chemistry / Q000235:genetics; D012471:Salivary Proteins and Peptides / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D000728:Androgens; C486321:Crisp1 protein, mouse; D008562:Membrane Glycoproteins; D011944:Receptors, Androgen; D012471:Salivary Proteins and Peptides; D003545:Cysteine