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. 2017 Oct 2;216(10):3275–3290. doi: 10.1083/jcb.201702177

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© 2017 MacDonald and Piper

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Figure 5. — Gtr1/Gtr2 control of EE>PM recycling is mediated through Vam6. (A) Model for how Gtr1 (GTP-bound)/Gtr2 (GDP-bound) heterodimer localizes to the vacuolar membrane via the Ego complex to activate TORC1. Gtr1 is activated by the GEF Vam6, which senses leucine via the leucyl-tRNA synthetase, Cdc60. Vam6, Gtr1, and Gtr2 are also required for recycling from early endosomes and work through Ltv1. (B and C) Localization of Ste3-GFP-DUb in the indicated mutant cells counter-labeled with FM4-64 for 60 min (B) or 2 min (C). Arrowheads indicate endosomal punta exhibiting colocalization. (D) Efflux of FM4-64 in wild-type cells and gtr1Δ, gtr2Δ, ltv1Δ, and vam6Δ cells carrying vector alone, or plasmids expressing Vam6, Gtr1, Gtr2, or Gtr1 and Gtr2 mutants locked in nucleotide-bound forms, Gtr1^GTP, Gtr1^GDP, Gtr2^GDP, or Gtr2^GTP. Bars, 5 µm.