Figure 2.

Interactions between Ufd2 and Rad23. (A) Two-hybrid interactions. Yeast cells were cotransformed with plasmids encoding Gal4 DNA-binding domain fused to Ufd2 protein indicated on the left with Gal4 activation domain fused to Rad23 or portions of Rad23, as indicated above the panel. Growth on -his plate is indicative of protein-protein interaction. (B) GST pull-down assays. The mixtures containing glutathione beads, various GST fusion proteins, and yeast extracts with overexpressed myc-tagged Ufd2 were incubated at 4°C for 2 h. Lanes 1 and 2 contain 5% input extracts with or without tagged Ufd2, respectively. The bound proteins were eluted, fractioned by SDS-PAGE, and immunoblotted with antibody to myc. Ufd2 and Rad23 have also been detected in a complex by a whole-genome approach (Ho et al., 2002).