Department of Molecular, Cell, and Cancer Biology; Program in Molecular Medicine
Biophysics | Cell Biology | Genomics | Molecular Biology | Structural Biology
Functional interactions between gene regulatory factors and chromatin architecture have been difficult to directly assess. Here, we use micrococcal nuclease (MNase) footprinting to probe the functions of two chromatin-remodeling complexes. By simultaneously quantifying alterations in small MNase footprints over the binding sites of 30 regulatory factors in mouse embryonic stem cells (ESCs), we provide evidence that esBAF and Mbd3/NuRD modulate the binding of several regulatory proteins. In addition, we find that nucleosome occupancy is reduced at specific loci in favor of subnucleosomes upon depletion of esBAF, including sites of histone H2A.Z localization. Consistent with these data, we demonstrate that esBAF is required for normal H2A.Z localization in ESCs, suggesting esBAF either stabilizes H2A.Z-containing nucleosomes or promotes subnucleosome to nucleosome conversion by facilitating H2A.Z deposition. Therefore, integrative examination of MNase footprints reveals insights into nucleosome dynamics and functional interactions between chromatin structure and key gene-regulatory factors.
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Citation: Cell Rep. 2015 Oct 6;13(1):61-9. doi: 10.1016/j.celrep.2015.08.071. Epub 2015 Sep 24. Link to article on publisher's site
DOI of Published Version
Hainer, Sarah J. and Fazzio, Thomas G., "Regulation of Nucleosome Architecture and Factor Binding Revealed by Nuclease Footprinting of the ESC Genome" (2015). Open Access Articles. 2593.
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