Figure 4. The C‐terminus of H2A is destabilized in the CENP‐A/CENP‐C^CR complex.

1. Graph showing the relative abundance of undigested DNA (145 bp) as a function of time during digestion with micrococcal nuclease (MNase) for the CENP‐A nucleosome (red), the CENP‐A/CENP‐C^CR complex (blue), and the CENP‐A/CENP‐C^motif complex (black).
2. Overlay of three cryo‐EM maps (gray, blue, and green) of the CENP‐A nucleosome obtained by sorting on the DNA entry/exit site (Fig EV2F). Distance between most open (gray) and most closed (green) map is 2 Å. The entry/exit site is boxed, and the corresponding model is shown on the right. Note that the density of the H2A C‐terminus is well defined.
3. Overlay of three cryo‐EM maps (gray, blue, and green) of the CENP‐A/CENP‐C^CR complex obtained by sorting on the DNA entry/exit site (Fig EV4F). The distance between the most open (gray) and most closed (green) maps is 9 Å. Map assigned to CENP‐C^CR is shown in magenta. The entry/exit site is boxed, and the corresponding model is shown on the right. Note the absence of a clearly defined density of the H2A C‐tail (indicated by the dotted circle).
4. Same type of data as in (A) for the CENP‐A nucleosome assembled with tailless H2A (H2A, 1–109) alone (pink) and in complex with CENP‐C^CR (light blue).
5. Schematic representation of the interplay between the N‐terminus of H3 or CENP‐A and the C‐terminus of H2A in regulating flexibility of nucleosomal DNA wrap. DNA (cyan); longer H3 αN (green); shorter CENP‐A αN (red); H2A C‐terminus (yellow); CENP‐C (magenta).

Data information: For (A, D), data are presented as mean (SD) for each time point based on three independent experiments and virtual gels are available in Fig EV5.