Coregulator exchange and sphingosine-sensitive cooperativity of steroidogenic factor-1, general control nonderepressed 5, p54, and p160 coactivators regulate cyclic adenosine 3',5'-monophosphate-dependent cytochrome P450c17 transcription rate.
Transcription of the cytochrome P450 17 (CYP17) gene is regulated by cAMP-dependent binding of steroidogenic factor-1 (SF-1) to its promoter in the adrenal cortex. Using temporal chromatin immunoprecipitation and mammalian two-hybrid experiments, we establish the reciprocal presence of coactivators [general control nonderepressed (GCN5), cAMP response element-binding protein-binding protein, p300, p300/cAMP response element-binding protein-binding protein CBP associated factor, p160s, polypyrimidine tract associated splicing factor, and p54(nrb)], corepressors (class I histone deacetylases, receptor interacting protein, nuclear receptor corepressor, and Sin3A), and SWI/SNF (human homolog of yeast mating type switching/sucrose nonfermenting) and imitation SWI chromatin remodeling ATPases on the CYP17 promoter during transcription cycles in the H295R adrenocortical cell line. A ternary GCN5/SRC-1/SF-1 complex forms on the CYP17 promoter with cAMP-dependence within 30 min of cAMP stimulation, and corresponds with SWI/SNF chromatin remodeling. This complex is sensitive to the SF-1 antagonist sphingosine and results in decreased transcription of CYP17. GCN5 acetyltransferase activity and carboxy terminus binding proteins alternatively mediate disassembly of the complex. This work establishes the temporal order of cAMP-induced events on the promoter of a key steroidogenic gene during SF-1-mediated transcription.
Dammer
Eric B
EB
School of Biology, Parker H. Petit Institute for Bioengineering & Biosciences, Georgia Institute of Technology, 310 Ferst Drive, Atlanta, Georgia 30332-0230, USA.
Leon
Adam
A
Sewer
Marion B
MB
D000107:Acetylation; D000595:Amino Acid Sequence; D003994:Bucladesine / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002460:Cell Line; D042002:Chromatin Assembly and Disassembly; D000242:Cyclic AMP / Q000378:metabolism*; D004257:DNA Polymerase II / Q000378:metabolism; D004268:DNA-Binding Proteins; D051548:Histone Acetyltransferases / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism*; D056920:Nuclear Receptor Coactivator 1; D050810:Octamer Transcription Factors / Q000378:metabolism*; D000072670:PTB-Associated Splicing Factor; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D016601:RNA-Binding Proteins / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D015398:Signal Transduction; D013110:Sphingosine / Q000378:metabolism*; D013254:Steroid 17-alpha-Hydroxylase / Q000235:genetics / Q000378:metabolism*; D054339:Steroidogenic Factor 1; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor
D003994:Bucladesine; D018797:Cell Cycle Proteins; D000242:Cyclic AMP; D004257:DNA Polymerase II; D004268:DNA-Binding Proteins; D051548:Histone Acetyltransferases; D018398:Homeodomain Proteins; D034521:Nuclear Matrix-Associated Proteins; D056920:Nuclear Receptor Coactivator 1; D050810:Octamer Transcription Factors; D000072670:PTB-Associated Splicing Factor; D016601:RNA-Binding Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D013110:Sphingosine; D013254:Steroid 17-alpha-Hydroxylase; D054339:Steroidogenic Factor 1; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C083214:NONO protein, human; C515057:NR5A1 protein, human; C538859:NCOA1 protein, human