High-Resolution Mapping of Sequence-Directed Nucleosome Positioning on Genomic DNA

Ross Fraser, D. Keszenman-Pereyra, Martin Simmen, J. Allan

Research output: Contribution to journalArticlepeer-review

Abstract

We have mapped in vitro nucleosome positioning on the sheep beta-lactoglobulin gene using high-throughput sequencing to characterise the DNA sequences recovered from reconstituted nucleosomes. This methodology surpasses previous approaches for coverage, accuracy and resolution and, most importantly, offers a simple yet rapid and relatively inexpensive method to characterise genomic DNA sequences in terms of nucleosome positioning capacity. We demonstrate an unambiguous correspondence between in vitro and in vivo nucleosome positioning around the promoter of the gene; identify discrete, sequence-specific nucleosomal structures above the level of the canonical core particle-a feature that has implications for regulatory protein access and higher-order chromatin packing; and reveal new insights into the involvement of periodically organised dinucleotide sequence motifs of the type GG and CC and not AA and TT, as determinants of nucleosome positioning-an observation that supports the idea that the core histone octamer can exploit different patterns of sequence organisation, or structural potential, in the DNA to bring about nucleosome positioning. (C) 2009 Elsevier Ltd. All rights reserved.
Original languageEnglish
Pages (from-to)292-305
Number of pages14
JournalJournal of Molecular Biology
Volume390
Issue number2
DOIs
Publication statusPublished - Jul 2009

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