FIGURE 4.
Interaction between Ypk2 and TORC2 is affected in avo3ts mutants. Tor2-Ypk2 or Avo1-Ypk2 interaction was determined by GST pulldown assays. A and C, WT and avo3ts cells expressing HA-Tor2 or Avo1-Myc were transformed with a plasmid expressing GST or GST-Ypk2. Exponentially growing cultures were incubated at indicated temperatures for 3 h and subjected to pulldown procedures. Pulled down proteins were analyzed by Western analysis using anti-HA or anti-Myc antibodies to detect HA-Tor2 or Avo1-Myc, whereas GST and GST-Ypk2 were visualized by Coomassie Blue staining. Expression levels of HA-Tor2 or Avo1-Myc in lysates are shown in the bottom panels. B and D, cells containing the empty vector (V), or an AVO1-, AVO2-, or AVO3-expressing plasmid as indicated were grown at 37 °C, and their lysates were subjected to the GST pulldown assay as in A and C. PD, pulldown.