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. 2010 Jul 16;11(8):612–618. doi: 10.1038/embor.2010.98

Figure 3.

Figure 3

The SCA7 domains of human ATXN7 and yeast Sgf73 bind to nucleosomes. (A) Tandem affinity purifications (TAP) of SAGA using TAP-tagged Spt20 from the indicated strains were analysed by western blot using PAP or HA antibody. (B) SCA7 domains from ATXN7, ATXN7L3 or Sgf73 fused to GST or GST alone were immobilized on glutathione-Sepharose beads and incubated with mononucleosomes. The retained material was analysed by Coomassie blue staining (Coo.) and western blotting using H2B or H3 antibodies, as indicated. All interactions were lost when the fusion proteins were pre-incubated with EDTA solution. (C) Schematic representation of ATXN7–SCA7 mutant domains used in GST-pulldown experiments. (D) Mononucleosomes retained by immobilized SCA7 domains were analysed by western blotting using H3 and H2B antibodies. GST, glutathione S-transferase; HA, haemagglutinin; PAP, peroxidase anti-peroxidase; SAGA, Spt–Ada–Gcn5 acetyltransferase; ZnF, zinc-finger.