Thimet oligopeptidase expression is differentially regulated in neuroendocrine and spermatid cell lines by transcription factor binding to SRY (sex-determining region Y), CAAT and CREB (cAMP-response-element-binding protein) promoter consensus sequences

Abstract

The zinc metalloprotease thimet oligopeptidase (EP24.15) is found predominantly in the neuroendocrine-gonadal axis where it is implicated in the processing of bioactive peptides, including GnRH (gonadotropin-releasing hormone), beta-neoendorphin, alpha-neoendorphin and dynorphin(1-8), the progression of spermatogenesis and the normal clearance of beta-amyloid in brain cells. Regulation of the enzyme's activity may occur in part by phosphorylation and redox disruption of intermolecular disulphide bridges. The elevated levels of both EP24.15 activity and mRNA within testicular and neuroendocrine tissues indicate that EP24.15 gene expression is differentially regulated. In the present paper, we present a detailed analysis of the rat EP24.15 promoter region previously isolated and partially characterized in this laboratory. Employing site-directed mutagenesis to create a series of promoter deletions and full-length promoter mutants, and measuring their activity in luciferase reporter gene and electrophoretic mobility-shift assays, we have shown that the transcription of the EP24.15 gene is differentially regulated in neuroendocrine and spermatid cell lines by transcription factor binding to SRY (sex-determining region Y), CAAT and CREB (cAMP-response-element-binding protein) promoter consensus sequences. The key to identifying the in vivo role of thimet oligopeptidase is likely to be found within the mechanisms by which it is regulated, and it is therefore of particular significance that EP24.15 expression is regulated by SRY and CREB/CREM (cAMP-response element modulator), the principle testes-determining protein and the major orchestrator of spermatogenesis respectively.