A C-terminal motif contributes to the plasma membrane localization of Arabidopsis STP transporters.

Membrane trafficking is highly organized to maintain cellular homeostasis in any organisms. Membrane-embedded transporters are targeted to various organelles to execute appropriate partition and allocation of their substrates, such as ions or sugars. To ensure the fidelity of targeting and sorting, membrane proteins including transporters have sorting signals that specify the subcellular destination and the trafficking pathway by which the destination is to be reached. Here, we have identified a novel sorting signal (called the tri-aromatic motif) which contains three aromatic residues, two tryptophans and one histidine, for the plasma membrane localization of sugar transporters in the STP family in Arabidopsis. We firstly found that a C-terminal deletion disrupted the sugar uptake activity of STP1 in yeast cells. Additional deletion and mutation analyses demonstrated that the three aromatic residues in the C-terminus, conserved among all Arabidopsis STP transporters, were critical for sugar uptake by not only STP1 but also another STP transporter STP13. We observed that, when the tri-aromatic motif was mutated, STP1 was largely localized at the endomembrane compartments in yeast cells, indicating that this improper subcellular localization led to the loss of sugar absorption. Importantly, our further analyses uncovered that mutations of the tri-aromatic motif resulted in the endoplasmic reticulum (ER) retention of STP1 and STP13 in plant cells, suggesting that this motif is involved at the step of ER exit of STP transporters to facilitate their plasma membrane localization. Together, we here identified a novel ER export signal, and showed that appropriate sorting via the tri-aromatic motif is important for sugar absorption by STP transporters.