FIG. 5.
An AHC1 mutant specifically affects the ADA HAT complex. Whole-cell extracts from a wild-type strain and a yeast strain bearing a mutation in AHC1 were partially purified with Ni2+ NTA agarose and Mono Q chromatography. (A) In the top panel, a typical fluorogram from a nucleosomal HAT assay with fractions from a Mono Q column prepared from a wild-type strain is presented. The four HAT complexes are indicated at the top. The lower panels show results from Western blotting with antibodies (α) raised against Ada2 and Gcn5. (B) Fluorogram from a nucleosomal HAT assay of Mono Q fractions prepared from the ahc1Δ yeast strain. NuA4, NuA3, and SAGA were eluted in the same fractions as in the wild type. ADA was absent in the AHC1 mutant. Western blots (lower panels) demonstrate that anti-Ada2 and anti-Gcn5 antibodies showed immunoreactivity only for SAGA (fractions 38 to 40).