Figure 6.

The dNLP-ISWI system can assemble containing histone variants H3. 3 and H2A.V. A, purification of Drosophila core histones with the H3.3 and H2A.V variants. The core histone proteins were synthesized in E. coli, purified, and analyzed by 15% polyacrylamide-SDS gel electrophoresis and staining with Coomassie Brilliant Blue R-250. The positions of the individual histones are shown. B, DNA supercoiling analysis of chromatin assembled with histones H3.3 and H2A.V. Chromatin assembly reactions were performed as described in the legend for Fig. 1B except that the H3.3 and H2A.V variants were used instead of the S phase–regulated H3 and H2A histones. The positions of nicked DNA (N), relaxed DNA (R), and supercoiled DNA (S) are shown. The black dot corresponds to a minor unknown contaminant. C, partial MNase digestion analysis of chromatin assembled with histones H3.3 and H2A.V. The samples were analyzed as described in the legend for Fig. 1C.