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. 2018 Jun 14;46(14):7138–7152. doi: 10.1093/nar/gky507

Figure 8.

Figure 8.

APLFAD functions as histone chaperone and prevents H2A-H2B-mediated DNA precipitation. (A) Native PAGE analysis of 167 base-pair (bp) DNA (1 μM) with the 601 nucleosome positioning sequence in presence or absence of APLFAD WT or mutants and H2A-H2B. Chaperone assay performed at 37°C. Lane 1: 50 bp DNA ladder (M). Lane 2: free DNA. Lane 3: DNA upon addition of 15 μM H2A-H2B. Lanes 4–15: DNA upon addition of 15 μM H2A-H2B preincubated with increasing concentrations (15, 45 or 90 μM) of APLFAD WT, Y476A, W485A or Y476A/W485A. (B) Calorimetric titration of APLFAD WT or mutants to H2A-H2B via ITC at 25°C. The resulting binding isotherms (upper panel) were fit to a one-set-of-sites binding mode. Best-fit values and fitting errors are shown in the table together with the derived thermodynamic parameters (lower panel). All data are obtained in 25 mM NaPi, pH 7.0, 300 mM NaCl.