Sp proteins play a critical role in histone deacetylase inhibitor-mediated derepression of CYP46A1 gene transcription

Maria João Nunes, Inês Milagre, Michael Schnekenburger, Maria João Gama, Marc Diederich, Elsa Rodrigues*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

37 Citations (Scopus)

Abstract

We investigated whether the CYP46A1 gene, a neuronal-specific cytochrome P450, responsible for the majority of brain cholesterol turnover, is subject to transcriptional modulation through modifications in histone acetylation. We demonstrated that inhibition of histone deacetylase activity by trichostatin A (TSA), valproic acid and sodium butyrate caused a potent induction of both CYP46A1 promoter activity and endogenous expression. Silencing of Sp transcription factors through specific small interfering RNAs, or impairing Sp binding to the proximal promoter, by site-directed mutagenesis, led to a significant decrease in TSA-mediated induction of CYP46A1 expression/promoter activity. Electrophoretic mobility shift assay, DNA affinity precipitation assays and chromatin immunoprecipitation assays were used to determine the multiprotein complex recruited to the CYP46A1 promoter, upon TSA treatment. Our data showed that a decrease in Sp3 binding at particular responsive elements, can shift the Sp1/Sp3/Sp4 ratio, and favor the detachment of histone deacetylase (HDAC) 1 and HDAC2 and the recruitment of p300/CBP. Moreover, we observed a dynamic change in the chromatin structure upon TSA treatment, characterized by an increase in the local recruitment of euchromatic markers and RNA polymerase II. Our results show the critical participation of an epigenetic program in the control of CYP46A1 gene transcription, and suggest that brain cholesterol catabolism may be affected upon treatment with HDAC inhibitors.

Original languageEnglish
Pages (from-to)418-431
Number of pages14
JournalJournal of Neurochemistry
Volume113
Issue number2
DOIs
Publication statusPublished - Apr 2010

Keywords

  • 24(S)-hydroxycholesterol
  • Brain cholesterol hom-eostasis
  • CYP46A1
  • Epigenetics
  • Histone acetylation
  • Sp transcription factors
  • Trichostatin A

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