Steroidogenic factor-1 contributes to the cyclic-adenosine monophosphate down-regulation of human SRY gene expression.
In mammals, male sex determination is initiated by SRY (sex-determining region of the Y chromosome) gene expression and followed by testicular development. This study describes specific down-regulation of the human SRY gene transcription by cAMP stimulation using reverse transcription-polymerase chain reaction experiments. Using transfection experiments, conserved nuclear hormone receptor (NHR1) and Sp1 consensus binding sites were identified as essential for this cAMP transcriptional response. Steroidogenic factor-1 (SF-1), a component of the sex-determination cascade, binds specifically to the NHR1 site and activates the SRY promoter. Activation of SF-1 was abolished by cAMP pretreatment of the cells, suggesting a possible effect of cAMP on the SF-1 protein itself. Indeed, human SF-1 protein contains at least two in vitro cAMP-dependent protein kinase (PKA) phosphorylation sites, leading after phosphorylation to a modification of both DNA-binding activity and interaction with general transcription factors such as Sp1. Taken together, these data suggest that cAMP responsiveness of human SRY promoter involves both SF-1 and Sp1 sites and could act via PKA phosphorylation of the SF-1 protein itself.
de Santa Barbara
P
P
Human Molecular Genetics Group, Institut de Génétique Humaine, CNRS UPR1142, 34396 Montpellier Cedex 5, France.
Méjean
C
C
Moniot
B
B
Malclès
M H
MH
Berta
P
P
Boizet-Bonhoure
B
B
D000242:Cyclic AMP / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000032:analysis; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D015536:Down-Regulation; D004591:Electrophoresis, Polyacrylamide Gel; D051717:Fushi Tarazu Transcription Factors; D018507:Gene Expression Regulation, Developmental; D006367:HeLa Cells; D018398:Homeodomain Proteins; D006801:Humans; D009687:Nuclear Proteins; D010766:Phosphorylation; D010957:Plasmids; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D020239:Receptor Cross-Talk / Q000502:physiology; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051876:Sex-Determining Region Y Protein; D016329:Sp1 Transcription Factor / Q000378:metabolism; D054339:Steroidogenic Factor 1; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology; D014162:Transfection
D017931:DNA Primers; D004268:DNA-Binding Proteins; D051717:Fushi Tarazu Transcription Factors; D018398:Homeodomain Proteins; C515057:NR5A1 protein, human; D009687:Nuclear Proteins; D018160:Receptors, Cytoplasmic and Nuclear; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein; D016329:Sp1 Transcription Factor; D054339:Steroidogenic Factor 1; D014157:Transcription Factors; D012313:RNA; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases