Chicken FANCD2 L234R exhibits impaired histone chaperone activity.

<p>(A) Schematic diagram of the supercoiling assay for nucleosome formation. Core histones were assembled on the relaxed plasmid DNA by a histone chaperone in the presence of wheat germ topoisomerase I. After deproteinization, the topoisomers were separated by agarose gel electrophoresis. (B) Supercoiling assays for nucleosome formation were performed with wild type (WT) cFANCD2 (lanes 4–7) and cFANCD2 L234R (lanes 9–12). Protein concentrations were 0 (lanes 3 and 8), 0.6 (lanes 4 and 9), 1.2 (lanes 5 and 10), and 1.8 µM (lanes 6, 7, 11, and 12). Highly supercoiled DNA is denoted as ‘sc DNA’. (C) Supercoiling assays for nucleosome formation were performed with WT cFANCD2 or cFANCD2 L234R, in the presence of WT cFANCI. The protein concentrations used in the assay are indicated at the top of the panel.