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J. Biol. Chem., Vol. 276, Issue 29, 27392-27399, July 20, 2001
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From the Transcriptional regulation of steroidogenic acute
regulatory protein (StAR) determines adrenal and gonadal cell
steroidogenesis. Chromatin immunoprecipitation assays were
combined with quantitative real-time polymerase chain reaction to
assess histone acetylation associated with the StAR promoter. MA-10
cells treated with 8-bromo-cAMP had increased acetylated histone H3
associated with the proximal (but not distal) StAR promoter, nascent
StAR transcripts, and progesterone production within 15 min, whereas
StAR mRNA increased at 30 min. At 360 min, steroidogenesis remained
elevated, but mRNA, nascent RNA, and StAR promoter-associated H3
acetylation all declined. StAR promoter-associated H4 acetylation was
unchanged by 8-bromo-cAMP treatment of MA-10 cells. In vivo
analysis of macaque and human granulosa cells showed that luteinization
was associated with increased StAR promoter-associated H3 acetylation. We conclude that acetylation of H3 (but not H4) associated with the
proximal promoter is associated with StAR gene transcription, that
chromatin modification occurs in discrete regions of the promoter, that
the initial steroidogenic response to 8-bromo-cAMP occurs prior to
increased StAR mRNA accumulation, and that MA-10 cell StAR gene
transcription and promoter-associated H3 acetylation are biphasic
during a 6-h treatment period. The union of the chromatin immunoprecipitation assay with quantitative real-time polymerase chain
reaction described and validated here should enhance the analysis of
gene expression.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AY007224.
Quantitative Analysis of the Hormone-induced
Hyperacetylation of Histone H3 Associated with the Steroidogenic Acute
Regulatory Protein Gene Promoter*
§,
Center for Research on Reproduction and
Women's Health, University of Pennsylvania, Philadelphia, Pennsylvania
19104-6142 and the ¶ Division of Reproductive Sciences, Oregon
Regional Primate Research Center, Beaverton, Oregon 97006
*
This work was supported by National Institutes of Health
Grants HD06274 (to J. F. S.), HD20869 (to R. L. S.), SCCPRR U54
HD18185 (Art Core), and RR00163.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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