FIGURE 9.

Edelfosine induces internalization of nutrient H⁺-symporters Can1p and Fur4p of the MCC, ubiquitination of Fur4p by edelfosine. Cells co-expressing Pma1p-RFP and either Can1p-GFP, Fur4p-GFP, or Fur4p-GFP-Dub were grown to log phase in the presence or absence of edelfosine (19 μm) for 1 h at 30 °C before imaging using confocal microscopy. a, Pma1p (MCP) and Can1p (MCC) are internalized upon treatment with edelfosine. BF, bright field. b, Pma1p (MCP) and Fur4p (MCC) are internalized upon treatment with edelfosine. c, fusion of the catalytic domain of Ubp7p (DUb) to the carboxyl-terminal end of Fur4p-GFP (53) prevents internalization of Fur4p, whereas Pma1p still gets internalized upon treatment with edelfosine. Arrows point to the co-localization of Pma1p and Fur4p-GFP-Dub in large patches at the PM induced by edelfosine. Such distribution is absent from control cells. d, quantitation of at least 100 cells from each condition. IC, intracellular; EDLF, edelfosine.