UVRAG cooperates with cargo receptors to assemble the ER‐phagy site

ER‐phagy is a selective autophagy process that targets specific regions of the endoplasmic reticulum (ER) for removal via lysosomal degradation. During cellular stress induced by starvation, cargo receptors concentrate at distinct ER‐phagy sites (ERPHS) to recruit core autophagy proteins and initiate ER‐phagy. However, the molecular mechanism responsible for ERPHS formation remains unclear. In our study, we discovered that the autophagy regulator UV radiation Resistance‐Associated Gene (UVRAG) plays a crucial role in orchestrating the assembly of ERPHS. Upon starvation, UVRAG localizes to ERPHS and interacts with specific ER‐phagy cargo receptors, such as FAM134B, ATL3, and RTN3L. UVRAG regulates the oligomerization of cargo receptors and facilitates the recruitment of Atg8 family proteins. Consequently, UVRAG promotes efficient ERPHS assembly and turnover of both ER sheets and tubules. Importantly, UVRAG‐mediated ER‐phagy contributes to the clearance of pathogenic proinsulin aggregates. Remarkably, the involvement of UVRAG in ER‐phagy initiation is independent of its canonical function as a subunit of class III phosphatidylinositol 3‐kinase complex II. Synopsis ER‐phagy initiation is localized to distinct sites on the ER, but the assembly mechanism of these ER‐phagy sites (ERPHS) remains unclear. This study identifies UVRAG as a novel regulator of ERPHS formation, operating independently to its canonical function in PI3KC3 complex II. UVRAG translocates to ERPHS and interacts with diverse ER‐phagy cargo receptors, such as FAM134B, ATL3, and RTN3L. UVRAG promotes oligomerization of ER‐phagy cargo receptors. UVRAG enhances the binding of ER‐phagy cargo receptors to Atg8 family proteins. UVRAG‐mediated ER‐phagy promotes the degradation of ER sheets, tubules, and aggregation‐prone ER luminal proteins, such as pathogenic proinsulin. The autophagy regulator UVRAG promotes degradation of specific regions of the endoplasmic reticulum independently of its role in PI3KC3 complex II.