Using chromatin immunoprecipitation to monitor 1alpha,25-dihydroxyvitamin D3-dependent chromatin activity on the human CYP24 promoter

The Journal of Steroid Biochemistry and Molecular Biology
Sami VäisänenCarsten Carlberg

Abstract

We applied the chromatin immunoprecipitation (ChIP) method for the analysis of 1alpha,25-dihydroxyvitamin D(3) (1,25-D(3))-dependent chromatin activity on the human 24-hydroxylase (CYP24) promoter in MCF-7 human breast cancer cells. In this pilot study we concentrated on the proximal promoter (+22 to -424) of the CYP24 gene, which includes the known 1,25-D(3) response element (VDRE) cluster. A constitutively active region of the human histone 4a gene (-40 to +285) served for normalization. Chromatin activity snapshots were taken 0, 30, 60, 120, 180, 240 and 300 min after the onset of stimulation with 1,25-D(3) and anti-acetylated histone 4 antibodies were used for ChIP. Our results suggest that ChIP is suitable for monitoring 1,25-D(3)-dependent changes of chromatin organization and can be used to reveal information about chromatin activity in living cells.

References

Jul 23, 1998·Critical Reviews in Eukaryotic Gene Expression·C Carlberg, P Polly
Jan 10, 2003·Journal of Cellular Biochemistry·Carsten Carlberg

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Citations

May 3, 2005·Nucleic Acids Research·Lasse SinkkonenCarsten Carlberg
Mar 7, 2007·Molecular and Cellular Biology·Maren EckeyAria Baniahmad
Mar 21, 2007·The Journal of Steroid Biochemistry and Molecular Biology·Ivy ChungDonald L Trump
Apr 30, 2005·The Journal of Steroid Biochemistry and Molecular Biology·Aria Baniahmad
Feb 17, 2007·Drug Metabolism and Disposition : the Biological Fate of Chemicals·A'edah Abu-BakarJukka Hakkola

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