Figure 5.

Effects of rapamycin on Rtg3p and Mks1p. (A) Effects of rapamycin and glutamate on the subcellular localization of Rtg3p-GFP and Mks1p-GFP. PSY142 ρ⁺ cells expressing Rtg3p-GFP or Mks1p-GFP from constructs transplaced into the respective chromosomal RTG3 or MKS1 loci treated for 30 min with 200 ng/ml rapamycin (+) or vehicle alone (−) were grown in YNBD with or without the addition of 0.2% glutamate. (B) Rapamycin induces dephosphorylation of Rtg3p and Mks1p. Extracts from PSY142 ρ⁺ cells grown in YNBD supplemented with 0.01% glutamate and treated with rapamycin or vehicle as described in A were analyzed by Western blotting with Rtg3p polyclonal anti-Rtg3p antiserum or monoclonal anti-myc antibody. Lanes 3 and 4 are extracts prepared from rtg3Δ cells (top) or from cells not containing the construct encoding Mks1(myc)₃ (bottom).