Plasma membrane to vacuole traffic induced by glucose starvation requires Gga2‐dependent sorting at the trans‐Golgi network

Background Information In the yeast Saccharomyces cerevisiae, acute glucose starvation induces rapid endocytosis followed by vacuolar degradation of many plasma membrane proteins. This process is essential for cell viability, but the regulatory mechanisms that control it remain poorly understood. Under normal growth conditions, a major regulatory decision for endocytic cargo occurs at the trans‐Golgi network (TGN) where proteins can recycle back to the plasma membrane or can be recognized by TGN‐localised clathrin adaptors that direct them towards the vacuole. However, glucose starvation reduces recycling and alters the localization and post‐translational modification of TGN‐localised clathrin adaptors. This raises the possibility that during glucose starvation endocytosed proteins are routed to the vacuole by a novel mechanism that bypasses the TGN or does not require TGN‐localised clathrin adaptors. Results Here, we investigate the role of TGN‐localised clathrin adaptors in the traffic of several amino acid permeases, including Can1, during glucose starvation. We find that Can1 transits through the TGN after endocytosis in both starved and normal conditions. Can1 and other amino acid permeases require TGN‐localised clathrin adaptors for maximal delivery to the vacuole. Furthermore, these permeases are actively sorted to the vacuole, because ectopically forced de‐ubiquitination at the TGN results in the recycling of the Tat1 permase in starved cells. Finally, we report that the Mup1 permease requires the clathrin adaptor Gga2 for vacuolar delivery. In contrast, the clathrin adaptor protein complex AP‐1 plays a minor role, potentially in retaining permeases in the TGN, but it is otherwise dispensable for vacuolar delivery. Conclusions and significance This work elucidates one membrane trafficking pathway needed for yeast to respond to acute glucose starvation. It also reveals the functions of TGNlocalised clathrin adaptors in this process. Our results indicate that the same machinery is needed for vacuolar protein sorting at the GN in glucose starved cells as is needed in the presence of glucose. In addition, our findings provide further support for the model that the TGN is a transit point for many endocytosed proteins, and that Gga2 and AP‐1 function in distinct pathways at the TGN. Research Article: Glucose starvation in yeast causes endocytosis of many cell surface proteins and their subsequent delivery to the yeast lysosome or vacuole. Here, we show t