C5 conserved region of hydrophilic C-terminal part ofSaccharomyces cerevisiaeNha1 antiporter determines its requirement of Erv14 COPII cargo receptor for plasma-membrane targeting

Erv14, a conserved cargo receptor of COPII vesicles, helps the proper trafficking of many but not all transporters to the yeast plasma membrane, for example, three out of five alkali-metal-cation transporters inSaccharomyces cerevisiae. Among them, the Nha1 cation/proton antiporter, which participates in cell cation and pH homeostasis, is a large membrane protein (985 aa) possessing a long hydrophilic C-terminus (552 aa) containing six conserved regions (C1-C6) with unknown function. A short Nha1 version, lacking almost the entire C-terminus, still binds to Erv14 but does not need it to be targeted to the plasma membrane. Comparing the localization and function ofScNha1 variants shortened at its C-terminus in cells with or without Erv14 reveals that onlyScNha1 versions possessing the complete C5 region are dependent on Erv14. In addition, our broad evolutionary conservation analysis of fungal Na+/H(+)antiporters identified new conserved regions in their C-termini, and our experiments newly show C5 and other, so far unknown, regions of the C-terminus, to be involved in the functionality and substrate specificity ofScNha1. Taken together, our results reveal that also relatively small hydrophilic parts of some yeast membrane proteins underlie their need to interact with the Erv14 cargo receptor.

Automated summary

The study demonstrates that some yeast membrane proteins, such as Nha1, rely on interacting with the Erv14 cargo receptor for proper trafficking to the plasma membrane. Specific variants of ScNha1 were found to be dependent on Erv14, shedding light on the functionality and substrate specificity of these proteins.