Figure 6: Arginine mutation at H4K5 and K12 causes mis-incorporation of the CENP-A–H4 complex into non-centromere regions.
From: Acetylation of histone H4 lysine 5 and 12 is required for CENP-A deposition into centromeres
(a) Representative images of SNAP-CENP-A labelled with TMR-Star in DT40 cells conditionally expressing H4_K5K12, H4_A5A12, and H4_R5R12. CENP-T was used as a centromere marker. CENP-A mis-incorporation was especially observed in cells expressing H4_R5R12. Bar, 10 μm. (b) Fluorescent intensity ratio of centromere signals for SNAP-CENP-A to non-centromere signals. When arginine mutant of H4 (H4_R5R12) was expressed, this ratio was lower than in cells expressing H4_K5K12 or H4_A5A12. Error bars represent s.d. Asterisk indicates statistically significance (P<0.0001) by Student's t-test. (N=100). (c) Increase of sequence reads in non-centromere region in cells expressing H4_R5R12 based on ChIP-seq analysis with anti-CENP-A antibody.
