FIGURE 7.

Grc3 PNK active site motifs do not disrupt interaction with Las1 in S. cerevisiae. (A,B) S. cerevisiae tetO₇-GRC3–Las1 5X Flag strains transformed with plasmids encoding Grc3 PNK variants were grown to mid-log phase at 30°C in the absence (A) and presence (B) of DOX (20 µg/mL). Cells were harvested, then lysed, and equal amounts of whole-cell lysate (WCL) were separated by SDS-PAGE and analyzed by western blot with an anti-Myc antibody (Grc3), an anti-Flag antibody (Las1), and an antitubulin antibody (loading control). Cells grown in the presence of DOX were further analyzed by coimmunoprecipitation with anti-Myc resin. Immunoprecipitated proteins were separated by SDS-PAGE and analyzed by western blot using an anti-Myc antibody (Grc3) and an anti-Flag antibody (Las1).