Phosphorylation switches protein disulfide isomerase activity to maintain proteostasis and attenuate ER stress.

Accumulated unfolded proteins in the endoplasmic reticulum (ER) trigger the unfolded protein response (UPR) to increase ER protein folding capacity. ER proteostasis and UPR signaling need to be regulated in a precise and timely manner. Here, we identify phosphorylation of protein disulfide isomerase (PDI), one of the most abundant and critical folding catalysts in the ER, as an early event during ER stress. The secretory pathway kinase Fam20C phosphorylates Ser357 of PDI and responds rapidly to various ER stressors. Phosphorylation of Ser357 induces an open conformation of PDI and turns it from a "foldase" into a "holdase", which is critical for preventing protein misfolding in the ER. Phosphorylated PDI also binds to the lumenal domain of IRE1α, a major UPR signal transducer, and attenuates excessive IRE1α activity. Importantly, PDI‐S359A knock‐in mice display enhanced IRE1α activation and liver damage under acute ER stress. We conclude that the Fam20C‐PDI axis constitutes a post‐translational response to maintain ER proteostasis and plays a vital role in protecting against ER stress‐induced cell death. Synopsis: Endoplasmic reticulum (ER) proteostasis and unfolded protein response (UPR) signaling need to be regulated in a precise and timely manner. Here, phosphorylation of protein disulfide isomerase (PDI) by secretory pathway kinase Fam20C turns PDI activity from "foldase" into "holdase", thereby protecting mouse liver from ER stress‐induced cell death. Fam20C phosphorylates ER‐associated PDI on Serine 357 rapidly after ER stress.Ser357 phosphorylation induces a functional switch that transforms PDI from a 'foldase' into a 'holdase' chaperone.Phosphorylated PDI safeguards ER proteostasis and attenuates excessive activity of UPR transducer IRE1α.PDI phosphorylation is critical for preventing liver damage under ER stress in vivo. [ABSTRACT FROM AUTHOR]