title: Transcriptional regulation of the rat Müllerian inhibiting substance type II receptor in rodent Leydig cells.
abstract: Müllerian inhibiting substance (MIS) causes regression of the fetal Müllerian duct on binding a heteromeric complex of types I and II cell-surface receptors in the fetal urogenital ridge. The MIS type II receptor (MISRII), which provides specificity for MIS, is also expressed in the adult testis, ovary, and uterus. The rat MISRII promoter was cloned to study the molecular mechanisms underlying its temporal and cell-specific expression. The 1.6-kilobase (kb) promoter contained no recognizable TATA or CAAT box, but there was a consensus Sp1 site upstream of the transcription initiation site. Two binding sites for the orphan nuclear receptor steroidogenic factor-1 (SF-1) are occupied in vitro by using nuclear extracts from R2C cells, an MIS-responsive rat Leydig cell line that expresses endogenous MISRII, with differing affinities, indicating that the distal SF-1 site is bound more avidly than is the proximal SF-1 site. R2C cells transfected with MISRII promoter/luciferase reporter constructs show a 12-fold induction with the 1.6-kb fragment and deletion of sequences upstream of -282-bp lowered luciferase expression to one-third. Mutation of both SF-1 sites greatly inhibited luciferase expression, whereas mutation of either site alone resulted in continuing activation by endogenous SF-1, indicating redundancy. In vitro binding and transcriptional analyses suggest that a proximal potential Smad-responsive element and an uncharacterized element also contribute to activation of the MISRII gene. R2C cells and MISRII promoter regulation can now be used to uncover endogenous transcription factors responsible for receptor expression or repression.
authors.affiliation: Pediatric Surgical Research Laboratories, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA. teixeira@helix.mgh.harvard.edu
authors.forename: J
authors.initials: J
authors.lastname: Teixeira
authors.forename: D J
authors.initials: DJ
authors.lastname: Kehas
authors.forename: R
authors.initials: R
authors.lastname: Antun
authors.forename: P K
authors.initials: PK
authors.lastname: Donahoe
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000378:metabolism; D051717:Fushi Tarazu Transcription Factors; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D018398:Homeodomain Proteins; D007985:Leydig Cells / Q000166:cytology / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear; D018000:Receptors, Peptide / Q000235:genetics*; D018125:Receptors, Transforming Growth Factor beta; D017422:Sequence Analysis, DNA; D054339:Steroidogenic Factor 1; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D051717:Fushi Tarazu Transcription Factors; D018398:Homeodomain Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D018000:Receptors, Peptide; D018125:Receptors, Transforming Growth Factor beta; D054339:Steroidogenic Factor 1; D014157:Transcription Factors; C084621:anti-Mullerian hormone receptor; C515060:steroidogenic factor 1, rat; D008156:Luciferases