FIG. 6.

Yng1p is required for NuA3 HAT activity. (A) Wild-type (YJW540) and yng1Δ (YJW541) TAF30 affinity-purified NuA3, normalized for levels of Sas3HA₃ by Western blotting, were assayed for HAT activity on free HeLa histones. The fluorogram and Coomassie brilliant blue stain of an SDS-PAGE of these histones are shown. (B) NuA3 purified from strains YJW540 and YJW541, normalized for levels of HAT activity on free histones, was assayed for HAT activity on HeLa nucleosomes. (C) Yng1p mediates interaction of NuA3 with nucleosomal arrays. Biotinylated G5E4-5S nucleosome arrays were bound to paramagnetic beads (Dynabeads) coupled to streptavidin. NuA3 purified from either wild-type or yng1Δ strains was added to the array template where indicated. The template beads were concentrated with a magnet, and the presence of Sas3p in the input, supernatants, and beads was analyzed by Western blotting.