Cross talk of two Krupple transcription factors regulates expression of the ovine FSH receptor gene

Abstract

The follicle-stimulating hormone receptor (FSHR) in gonadal cells is required for normal folliculogenesis and spermatogenesis. To understand its regulation, we identified a CACC-box from −46 to −67 of the ovine FSHR promoter. Antibody supershift with a 22-bp DNA probe and nuclear extract from a Sertoli cell line demonstrated that a testis-specific zinc finger protein, ZNF202, might be one of the binding proteins. Western blots using ZNF202 antibody and Southwestern blot analyses with the DNA probe detected the same 60 kDa protein in both Sertoli and ovarian granulosa cell lines. Gel shift assays also revealed that the DNA–protein complex from Sertoli cells overexpressing the human Ras-responsive element binding protein-1 (RREB-1) migrated the same way as the complex containing endogenous CACC-box binding protein. Transfection studies indicated that ZNF202 repressed ovine FSHR promoter activity whereas RREB-1 was likely to function as an activator. These data suggest that selective expression and cross talk of functionally distinctive Krupple transcription factors could regulate tissue- and stage-specific expression of FSHR gene.