Figure 1. Rep8 interacts with p97 via the UBX domain.

(A) Yeast two-hybrid analyses of the p97-Rep8 interaction using the HIS3 reporter gene. Co-transformation of bait p97 with prey Rep8 supported cell growth under conditions selecting for the interaction (in the absence of histidine and presence of 25 mM 3-aminotriazol (3AT); right panel). The known p97-p47 interaction served as a control. (B) Schematic diagram of the Rep8 domain organization and the various truncations used in the precipitation experiments. (C) Purified p97 was incubated with the indicated, immobilized GST fusion proteins before analysis of bound proteins by SDS-PAGE and blotting using antibodies specific for p97 (top panel) or GST (lower panel).