Role of CACC-box in the regulation of ovine follicle-stimulating hormone receptor expression.
Tissue-specific and stage-specific expression of follicle-stimulating hormone receptor (FSH-R) in granulosa and Sertoli cells is required for normal development of ovarian follicles and germ cells. However, little is known of the transcription factors that regulate the FSH-R gene and its promoter. Using an ovine FSH-R promoter as a model system, we have identified a second DNase I footprinting 2 (FP2) region from -46 to -67 of the strongest ovine FSH-R promoter (-200 to +163) relative to the transcription start site. Electrophoretic mobility shift assay with a 22-base pair DNA probe (-46 to -67) and nuclear extracts from Sertoli (15P1) and granulosa (JC-410) cell lines demonstrated a sequence-specific DNA-protein complex. Further Southwestern and UV cross-linking analyses detected three predominant proteins of molecular weights 87, 60, and 50 kDa present in both Sertoli and granulosa cells bound to a 32P-labeled DNA probe as a complex. Gel competition experiments with DNA probes containing known Krupple-like factor binding sites revealed that the testis-specific zinc finger protein, ZNF202-like factor, Ras-responsive element binding protein-like factor, or both, may be among the potential candidate regulators. Mutation within the CACC box of the promoter abolished Krupple-like factor binding and significantly diminished promoter activity in both gonadal cells. These data suggest that Krupple-like transcription factors may play a role in the regulation of ovine FSH-R expression.
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Molecular Reproduction Research Laboratory, Clinical Research Institute of Montréal, Montréal, Québec, H2W 1R7, Canada.
Sairam
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D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D018983:DNA Footprinting; D004268:DNA-Binding Proteins; D003850:Deoxyribonuclease I; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D005786:Gene Expression Regulation; D051379:Mice; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011962:Receptors, FSH / Q000235:genetics*; D012756:Sheep; D014162:Transfection; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D011962:Receptors, FSH; C085482:Zbtb10 protein, rat; D003850:Deoxyribonuclease I