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. 2012 Oct 11;7(10):e47162. doi: 10.1371/journal.pone.0047162

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© 2012 Lo et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(a) Amino acid sequences for the wild-type H2A acidic patch (WT) and uncharged mutant (STT). Acidic residues are highlighted, and mutated residues are underlined. (b) MgCl₂ dependent oligomerization of wild type and H2A-STT containing chromatin. Chromatin was incubated with the indicated concentrations of MgCl₂ and centrifuged in a microfuge. Supernatants were electrophoresed on agarose gels and stained with SYBR gold; the % of the template remaining in the supernatant was determined by comparison with the 0 mM MgCl₂ control. (c) Summary of chromatin oligomerization assays. (d) Restriction enzyme accessibility (REA) assays on chromatin templates with 12 nucleosomes. The chromatin template contains a unique restriction site (HhaI) that is normally occluded by nucleosomes but is exposed upon Swi/Snf-mediated chromatin remodeling. The first two lanes are negative and positive controls (with or without Swi/Snf, no PSC) demonstrating that the HhaI site becomes more accessible in the presence of Swi/Snf. (e) Summary of REA assay on chromatin templates assembled with rec-WT and H2A-STT histones. Percent inhibition is calculated as .

Inline graphic — (a) Amino acid sequences for the wild-type H2A acidic patch (WT) and uncharged mutant (STT). Acidic residues are highlighted, and mutated residues are underlined. (b) MgCl₂ dependent oligomerization of wild type and H2A-STT containing chromatin. Chromatin was incubated with the indicated concentrations of MgCl₂ and centrifuged in a microfuge. Supernatants were electrophoresed on agarose gels and stained with SYBR gold; the % of the template remaining in the supernatant was determined by comparison with the 0 mM MgCl₂ control. (c) Summary of chromatin oligomerization assays. (d) Restriction enzyme accessibility (REA) assays on chromatin templates with 12 nucleosomes. The chromatin template contains a unique restriction site (HhaI) that is normally occluded by nucleosomes but is exposed upon Swi/Snf-mediated chromatin remodeling. The first two lanes are negative and positive controls (with or without Swi/Snf, no PSC) demonstrating that the HhaI site becomes more accessible in the presence of Swi/Snf. (e) Summary of REA assay on chromatin templates assembled with rec-WT and H2A-STT histones. Percent inhibition is calculated as .