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. 2015 Jun 5;4:e06249. doi: 10.7554/eLife.06249

Figure 2. A bound Gal4DBD affects the directionality of SWI/SNF remodeling and ISW1a remodeling differently.

Nucleosomes were remodeled by either 1 nM ISW1a or 1 nM SWI/SNF with 1 mM ATP for 1 min, a time sufficiently short that the majority of nucleosomes were not remodeled (Figure 2—figure supplement 2). Each DNA template was subsequently unzipped. For templates used in (D)–(G), the 601NPE was separated from the Gal4 binding sequence by 10 bp. (A) Distribution of the location of a nucleosome before remodeling. Data were pooled from measurements on multiple nucleosomal DNA molecules. (B) Distribution of the location of a nucleosome remodeled by ISW1a in the absence of Gal4DBD. (C) Distribution of the location of a nucleosome remodeled by SWI/SNF in the absence of Gal4DBD. (D) Distribution of the location of a nucleosome remodeled by ISW1a with a bound Gal4DBD initially located upstream of the 601NPE. (E) Distribution of the location of a nucleosome remodeled by SWI/SNF with a bound Gal4DBD initially located upstream of the 601NPE. (F) Distribution of the location of the nucleosome remodeled by ISW1a with a bound Gal4DBD initially located downstream of the 601NPE. (G) Distribution of the location of a nucleosome remodeled by SWI/SNF with a bound Gal4DBD initially located downstream of the 601NPE.

DOI: http://dx.doi.org/10.7554/eLife.06249.008

Figure 2—source data 1. Comparison of unzipping force signatures of a nucleosome before and after remodeling.
We used unzipping to characterize the structure of a nucleosome before or after remodeling by either ISW1a or SWI/SNF, in the presence or absence of Gal4DBD. The structural features include the maximum force in the first force cluster, the maximum force in the second force cluster, the width of each cluster, and the distance between the two clusters. Errors show s.d.
DOI: http://dx.doi.org/10.7554/eLife.06249.009
elife06249s001.docx (17.6KB, docx)
DOI: 10.7554/eLife.06249.009

Figure 2.

Figure 2—figure supplement 1. Directionality of ISW1a and SWI/SNF remodeling of a nucleosome in close proximity to a bound Gal4DBD.

Figure 2—figure supplement 1.

The figure panels A to G show the raw traces for the corresponding panels in Figure 2.
DOI: http://dx.doi.org/10.7554/eLife.06249.010
Figure 2—figure supplement 2. Determination of fractions of remodeled nucleosome.

Figure 2—figure supplement 2.

(A) For each nucleosome distribution, remodeled by either ISW1a or SWI/SNF as shown in Figure 2B,C, was fit to a double Gaussian function. One Gaussian (narrow green curve) corresponds to the distribution of unremodeled nucleosomes and the other (broader green curve) to the distribution of remodeled nucleosomes. The sum of the two Gaussians is shown as the red curve. (B) A table summarizing fractions of unremodeled percentage and remodeled percentage for both ISW1a and SWI/SNF.
DOI: http://dx.doi.org/10.7554/eLife.06249.011