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. 2012 Jul 17;1(9):884–888. doi: 10.1242/bio.20122022

Fig. 3. Both TORC1 and TORC2 regulate Rps6 phosphorylation while Maf1 phosphorylation is TORC1 specific.

Fig. 3.

All cells were grown in EMMG (A) Phosphorylation of Rps6 is TORC1 and TORC2 dependent. WT and temperature sensitive TORC1 mutants mip1.310 (Raptor) and tor2.51 were cultured to exponential phase at permissive temperature (28°C). The cells were heat shocked at 37°C for 2 hr (left). Deletion of tor1, ste20 (Rictor) or sin1 reduced the phosphorylation level of Rps6 (right). (B) Gad8 S546 phosphorylation is TORC2 specific. All strains were treated as in (A) and the phosphorylated protein was detected by P-Gad8.S546 specific antibodies and total Gad8 by anti-Gad8 antibodies. (C) Maf1 phosphorylation is TORC1 but not TORC2 dependent. TORC1 and TORC2 mutants expressing Maf1.pk were treated as in (A). Maf1.pk was detected by anti-pk antibodies. (D) Rapamycin inhibits Maf1.pk phosphorylation. Cells were grown and treated with rapamycin, and Maf1.pk was detected as previously described. (E) Gad8 is not required for Maf1 phosphorylation. (F) Slower migrating Maf1.pk bands represent phosphorylation. When λ phosphatase was added to immunoprecipitated Maf1.pk, only the faster migrating non-phosphorylated Maf1.pk was observed.