An ER-accumulated mutant of yeast Pma1 causes membrane-related stress to induce the unfolded protein response.

Upon dysfunction of the endoplasmic reticulum (ER), namely ER stress, eukaryotic cells provoke the unfolded protein response (UPR), which is triggered by ER stress sensors including Ire1. While the ER luminal domain of Ire1 is known to directly recognize misfolded soluble proteins accumulated in the ER, the transmembrane domain of Ire1 is involved in its self-association and activation upon membrane lipid-related abnormalities, which are so-called lipid bilayer stress (LBS). Here we inquired how the ER accumulation of misfolded transmembrane proteins induces the UPR. In yeast Saccharomyces cerevisiae cells, a multi-transmembrane protein, Pma1, is not transported to the cell surface but aggregates on the ER membrane when carrying a point mutation (Pma1-2308). Here, we show that GFP-tagged Ire1 co-localized with the Pma1-2308-mCherry puncta. This co-localization and the UPR induced by Pma1-2308-mCherry were compromised by a point mutation in Ire1 that specifically impairs its activation upon LBS. We presume that Pma1-2308-mCherry locally affects the properties (probably the thickness) of the ER membrane at its aggregation sites, where Ire1 is then recruited, self-associated, and then activated.