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Chromosomal mapping of core histone acetylation by immunoselection
C Crane-Robinson1, T R Hebbes, A L Clayton
1Biophysics Laboratories, University of Portsmouth, United Kingdom.
Methods (San Diego, Calif.)
|May 1, 1997
Summary
Histone acetylation, a marker of active genes, is a prerequisite for open chromatin structure, not a consequence of transcription. This modification may generate or maintain the open structure of poised and active genes.
Area of Science:
- Biochemistry
- Molecular Biology
- Epigenetics
Background:
- Histone acetylation marks active genes.
- The spatial and temporal distribution of this modification is not well understood.
Purpose of the Study:
- To investigate the distribution of histone acetylation.
- To determine if acetylation precedes or follows gene activation.
Main Methods:
- Used affinity-purified antibodies against acetylated lysine.
- Employed immunoselection with mononucleosomes and chromatin fragments.
- Slot-blotting and gene-specific probes to assess acetylation levels.
Main Results:
- Elevated acetylation levels were detected on histones associated with specific gene sequences.
- Acetylation uniformly covered the beta-globin locus in chicken erythrocytes, correlating with open chromatin.
- The inactive but poised PDGF-beta gene was hyperacetylated, even before induction.
Conclusions:
- Histone acetylation is a prerequisite for, not a consequence of, transcription.
- Acetylation may generate or maintain the open chromatin structure of poised and active genes.