FIG 2.

BS3 cross-linking of UAF and Uaf30-less subcomplex. (A) Coomassie blue-stained SDS-PAGE gel of purified holo-UAF (right) and Uaf30-less UAF (left) used in our cross-linking experiments. (B) Chemical cross-linking of recombinant holo-UAF. Products of incubation of the increasing concentrations from 0.2 to 10 mM BS3 are shown, visualized by SDS-PAGE and silver staining. (C to G) Western blot analysis of BS3 cross-linked Uaf30-less UAF. Recombinant UAF lacking Uaf30 was either left untreated or treated with increasing concentrations of BS3 cross-linker (0.2 to 4 mM) and then analyzed by Western blotting with various UAF subunit antibodies. Antibodies included histone H3 (C), histone H4 (D), Rrn10 (E), Rrn5 (F), and Rrn9 (G). Predicted UAF cross-linked intermediates, based on their calculated molecular weights, are labeled on the right of each Western blot image.