Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cyclic AMP / physiology*
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Cyclic AMP-Dependent Protein Kinases / analysis
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DNA Primers / chemistry
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DNA-Binding Proteins / biosynthesis
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DNA-Binding Proteins / genetics*
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DNA-Binding Proteins / metabolism
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DNA-Binding Proteins / physiology*
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Down-Regulation
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Electrophoresis, Polyacrylamide Gel
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Fushi Tarazu Transcription Factors
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Gene Expression Regulation, Developmental*
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HeLa Cells
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Homeodomain Proteins
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Humans
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Nuclear Proteins*
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Phosphorylation
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Plasmids
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Precipitin Tests
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Promoter Regions, Genetic
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Protein Processing, Post-Translational
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RNA / chemistry
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RNA / isolation & purification
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Receptor Cross-Talk / physiology
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Receptors, Cytoplasmic and Nuclear / genetics
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Receptors, Cytoplasmic and Nuclear / physiology
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Reverse Transcriptase Polymerase Chain Reaction
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Sex-Determining Region Y Protein
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Sp1 Transcription Factor / metabolism
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Steroidogenic Factor 1
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Transcription Factors / physiology*
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Transcription, Genetic / physiology
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Transfection
Substances
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DNA Primers
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DNA-Binding Proteins
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Fushi Tarazu Transcription Factors
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Homeodomain Proteins
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NR5A1 protein, human
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Nuclear Proteins
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Receptors, Cytoplasmic and Nuclear
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SRY protein, human
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Sex-Determining Region Y Protein
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Sp1 Transcription Factor
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Steroidogenic Factor 1
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Transcription Factors
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RNA
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Cyclic AMP
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Cyclic AMP-Dependent Protein Kinases