Fig. 6: The Dsup HMGN-like motif and C-terminal region each contribute to nucleosome binding. | Nature Communications

Fig. 6: The Dsup HMGN-like motif and C-terminal region each contribute to nucleosome binding.

From: Multivalent binding of the tardigrade Dsup protein to chromatin promotes yeast survival and longevity upon exposure to oxidative damage

Fig. 6

a Impact of ionic strength (150 mM (normal saline, 0.9%) or 250 mM NaCl) on binding of Dsup (WT: 1 - 0 μM in two-fold serial dilutions) to unmodified nucleosome (rNuc on 147 bp DNA; 10 nM) or free DNA (147 x 601: 2.5 nM). b salDNA (1μg/ml in 150 mM NaCl) was a more effective competitor of Dsup binding to free-DNA vs. to nucleosomes, suggesting the latter involves multivalent engagement. c Mutation of the Dsup HMGN-like motif (aa 363-370; charge reversing -3R/3E, or charge neutralizing -8A) or deletion of the adjacent C-terminal region (Δ371-455; ΔC) each compromised DNA / nucleosome binding compared to Dsup (WT) (see also Supplementary Fig. 8). Note the relative binding of each allele to free DNA or nucleosome targets, where Dsup HMGN mutants preferred free DNA while Dsup ΔC preferred nucleosomes. Key and reaction conditions as in (b). Data are the average and standard deviation of two technical repeats. Source data are provided as a Source Data file.

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