Endogenous expression of Müllerian inhibiting substance in early postnatal rat sertoli cells requires multiple steroidogenic factor-1 and GATA-4-binding sites.
Müllerian inhibiting substance (MIS) is a key element required to complete mammalian male sex differentiation. The expression pattern of MIS is tightly regulated in fetal, neonatal, and prepubertal testes and adult ovaries and is well conserved among mammalian species. Although several factors have been shown to be essential to MIS expression, its regulatory mechanisms are not fully understood. We have examined MIS promoter activity in 2-day postnatal primary cultures of rat Sertoli cells that continue to express endogenous MIS mRNA. Using this system, we found that the region between human MIS-269 and -192 is necessary for full MIS promoter activity. We identified by DNase I footprint and electrophoretic mobility-shift analyses a distal steroidogenic factor-1 (SF-1)-binding site that is essential for full promoter activity. Mutational analysis of this new distal SF-1 site and the previously identified proximal SF-1 site showed that both are necessary for transcriptional activation. Moreover, the proximal promoter also contains multiple GATA-4-binding sites that are essential for functional promoter activity. Thus multiple SF-1- and GATA-4-binding sites in the MIS promoter are required for normal tissue-specific and developmental expression of MIS.
Watanabe
K
K
Pediatric Surgical Research Laboratories and Gene Therapy Center, Massachusetts General Hospital, Department of Surgery, Harvard Medical School, Boston, MA 02114, USA.
Clarke
T R
TR
Lane
A H
AH
Wang
X
X
Donahoe
P K
PK
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D054304:Anti-Mullerian Hormone; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000378:metabolism*; D051717:Fushi Tarazu Transcription Factors; D050991:GATA4 Transcription Factor; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D017930:Genes, Reporter; D006023:Glycoproteins; D006131:Growth Inhibitors / Q000235:genetics*; D018398:Homeodomain Proteins; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear; D012708:Sertoli Cells; D054339:Steroidogenic Factor 1; D013735:Testicular Hormones / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation; D014162:Transfection
D004268:DNA-Binding Proteins; D051717:Fushi Tarazu Transcription Factors; D050991:GATA4 Transcription Factor; D006023:Glycoproteins; D006131:Growth Inhibitors; D018398:Homeodomain Proteins; C515057:NR5A1 protein, human; D009687:Nuclear Proteins; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D054339:Steroidogenic Factor 1; D013735:Testicular Hormones; D014157:Transcription Factors; C515060:steroidogenic factor 1, rat; D054304:Anti-Mullerian Hormone