Candidate Sertoli cell specific promoter element for a TGFβ family member (Amh) and a 3′ UTR enhancer/repressor for the same gene

Abstract

Comparison between 400 nt of mouse DNA sequence immediately upstream (5′) of the coding sequence of the Sertoli cell expressed genes, Amh and Tsx, identified a 33 nt sequence with a significant identity: this was considered to be a candidate Sertoli specific regulatory element (SSRE). Another highly conserved sequence has been identified immediately downstream (3′) of the Amh polyadenylation signal (DSRE). The action and specificity of these two putative regulatory elements, inserted into a reporter vector (pd2EGFP), has been investigated by transient transfection of mouse fibroblast and Sertoli cells. When combined together or alone, with a minimal thymidine kinase (Tk) promoter, SSRE showed a weak incremental action on the expression of the reporter gene, regardless of the cell type: in contrast there was a 2- to 3-fold decrease when DSRE was present. However, in the absence of Tk there was evidence for a strong synergy between SSRE and DSRE, which was significantly greater in the Sertoli as compared with the fibroblast cells. These results support the view that SSRE exhibits a degree of Sertoli specificity and acts synergistically with DSRE in controlling the expression of Amh.