Nucleic Acids Research, 1982, Vol. 10, No. 6 2017-2042
© 1982
MOLECULAR BIOLOGY |
Properties of active nucleosomes as revealed by HMG 14 and 17 chromatography
Laboratory of Molecular Biology, Medical Research Council Centre Hills Road, Cambridge CB2 2QH, UK
Received December 1, 1981. Revised February 10, 1982. Accepted February 22, 1982.
Nucleosomes fran actively transcribed genes (active nucleosomes) contain nonhistone proteins HMG 14 and 17 and are preferentially sensitive to digestion by DNAse I. Active nucleosomes isolated by chromatography on an HMG 14 and 17 glass bead affinity column were analyzed with respect to overall structure, accessory nonhistone components and modifications to the ENA and histones. the experiments lead to the following conclusions: The DNR in the active nucleosome is undermethylated compared to bulk DNA; topoisomerase I is a non-stoichiometric component of the active nucleosomes fraction; the level of histone acetylation is enriched in active nucleosames, but the extent of enrichment cannot account for HMG binding; and the two histone H3 molecules in the active nucleosome can dimerize more readily and are, therefore, probably closer together than those in the bulk of the nucleosomes. Additionally it is shown that HMG 14 and 17 prefer to bind to single- vs. double-stranded nucleic acids. The role of HMG 14 and 17 in producing a highly DNAse I sensitive structure and correspondingly helping to facilitate transcription is discussed in terms of these properties.
*Present address: Cold Spring Harbor Laboratory, P.O. Box 100, Cold Spring Harbor, NY 11724, USA
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