Figure 5.
The acetylation kinetics of nucleosomes at H3K9 by ATACHAT and SAGAHAT. Acetylation assay by the ATACHAT (A) or SAGAHAT (B) module at increasing time with and without Gal4–VP16 at H3K9ac. The fluorescence of Cy5-labeled histone H2A within the nucleosome was used to observe nucleosome loading. Acetylation efficiency was tested by western blot using antibodies recognizing histone H3K9ac. Images are representative of three independent experiments with similar results (n = 3). Error bars represent standard error of the mean. (C) Graph of panel (A). (D) Graph of panel (B). The time courses were fit to a single exponential as described in the ‘Materials and methods’ section to determine characteristic time, τ, for the acetylation reaction.