Chromatin Structure Analysis of the Mouse Xist Locus

The Xist gene is expressed exclusively from the inactive X chromosome and plays a central role in regulating X chromosome inactivation. Here we describe experiments aimed at defining the extent of the active chromatin domain of the expressed Xist allele. By using an allele-specific general DNaseI se...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1999-06, Vol.96 (13), p.7155-7160
Main Authors: McCabe, Veronica, Formstone, Emma J., O'Neill, Laura P., Turner, Bryan M., Brockdorff, Neil
Format: Article
Language:English
Subjects:
Acetylation
Alleles
Animals
Biochemistry
Biological Sciences
Cells
Chromatin
Chromatin - chemistry
Chromatin - genetics
Chromatin - metabolism
Genes
Genetic loci
Histones
Mice
RNA, Long Noncoding
RNA, Untranslated
Rodents
Somatic cells
Space probes
Transcription Factors - genetics
X chromosome
X Chromosome - genetics
X Chromosome inactivation
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Summary:The Xist gene is expressed exclusively from the inactive X chromosome and plays a central role in regulating X chromosome inactivation. Here we describe experiments aimed at defining the extent of the active chromatin domain of the expressed Xist allele. By using an allele-specific general DNaseI sensitivity assay we show that there is preferential digestion of the expressed allele at sites within the transcribed locus but not in flanking sites located up to 70 kb 5′. A putative proximal boundary for the Xist domain is located within 10 kb upstream of promoter P1. Chromatin in the expressed domain was found to be acetylated at H4 in XX somatic cells but also in XY cells, where Xist is never expressed. A single clear exception to this was the Xist promoter, which is acetylated only in XX cells. These observations concur with the view that H4 acetylation may not be a general marker of active chromatin domains and further support data implicating local promoter acetylation as being of primary functional significance in vivo.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.96.13.7155