The acyltransferase Gpc1 is both a target and an effector of the unfolded protein response in Saccharomyces cerevisiae
Journal of Biological Chemistry
acyltransferase, glycerophosphocholine, Gpc1, Ire1, lipid remodeling, lysophosphatidylcholine, membrane bilayer stress, phosphatidylcholine, phospholipid metabolism, unfolded protein response, yeast
The unfolded protein response (UPR) is sensitive to proteotoxic and membrane bilayer stress, both of which are sensed by the ER protein Ire1. When activated, Ire1 splices HAC1 mRNA, producing a transcription factor that targets genes involved in proteostasis and lipid metabolism, among others. The major membrane lipid phosphatidylcholine (PC) is subject to phospholipase-mediated deacylation, producing glycerophosphocholine (GPC), followed by reacylation of GPC through the PC deacylation/reacylation pathway (PC-DRP). The reacylation events occur via a two-step process catalyzed first by the GPC acyltransferase Gpc1, followed by acylation of the lyso-PC molecule by Ale1. However, whether Gpc1 is critical for ER bilayer homeostasis is unclear. Using an improved method for C14-choline-GPC radiolabeling, we first show that loss of Gpc1 results in abrogation of PC synthesis through PC-DRP and that Gpc1 colocalizes with the ER. We then probe the role of Gpc1 as both a target and an effector of the UPR. Exposure to the UPR-inducing compounds tunicamycin, DTT, and canavanine results in a Hac1-dependent increase in GPC1 message. Further, cells lacking Gpc1 exhibit increased sensitivity to those proteotoxic stressors. Inositol limitation, known to induce the UPR via bilayer stress, also induces GPC1 expression. Finally, we show that loss of GPC1 induces the UPR. A gpc1Δ mutant displays upregulation of the UPR in strains expressing a mutant form of Ire1 that is unresponsive to unfolded proteins, indicating that bilayer stress is responsible for the observed upregulation. Collectively, our data indicate an important role for Gpc1 in yeast ER bilayer homeostasis.
Hrach, V., King, W., Nelson, L., Conklin, S., Pollock, J., & Patton-Vogt, J. (2023). The acyltransferase Gpc1 is both a target and an effector of the unfolded protein response in Saccharomyces cerevisiae. Journal of Biological Chemistry, 299 (7). https://doi.org/10.1016/j.jbc.2023.104884