Androgen induction of the SVS family related protein MSVSP99: identification of a functional androgen response...

The gene encoding MSVSP99 (mouse seminal vesicle secretory protein of 99 amino acids) is specifically expressed in the mouse seminal vesicle under androgenic control. To study hormonal regulation, fragments of the 5'-flanking region, extending from -2365 to +16 were linked to the chloramphenicol acetyl transferase (CAT) gene and cotransfected with an androgen receptor expression vector into CV-1 cells. A minimal region (-387 to +16) was sufficient for full androgen induction. Further deletion, up to nt-261, almost completely abolished androgen inducibility. DNase I footprinting and band-shift assays, using the DNA binding domain of the androgen receptor (AR-DBD), revealed three AR binding sites: two putative androgen response elements (AREs) occurring at positions -361 (AREd) and -208 (AREp), and an androgen receptor binding region (ARBR) located between positions -317 and -293. Transient transfection assays revealed that site-directed mutation in AREp abolished androgen induced expression, whereas mutation in AREd or in ARBR had no effect. The results demonstrate that AREp is a functional sequence that must cooperate with additional cis-acting elements, located between -387 and -261, for androgen induction of the MSVSP99 gene.