Coat protein complex II (COPII) proteins form vesicles from the endoplasmic reticulum to export cargo molecules to the Golgi apparatus. Among the many proteins involved in this process, Sec12 is a key regulator, functioning as the guanosine diphosphate (GDP) exchange factor for Sar1p, the small guanosine triphosphatase (GTPase) that initiates COPII assembly. Here we show that overexpression of phospholipase B3 in the thermosensitive sec12-4 mutant partially restores growth and protein transport at non-permissive temperatures. Lipidomics analyses of these cells show a higher content of lysophosphatidylinositol (lysoPI), consistent with the lipid specificity of PLB3. Furthermore, we show that lysoPI is specifically enriched in COPII vesicles isolated from in vitro budding assays. As these results suggested that lysophospholipids could facilitate budding under conditions of defective COPII coat dynamics, we reconstituted COPII binding onto giant liposomes with purified proteins and showed that lysoPI decreases membrane rigidity and enhances COPII recruitment to liposomes. Our results support a mechanical facilitation of COPII budding by lysophospholipids.

Accurate maintenance of organelle identity in the secretory pathway relies on retention and retrieval of resident proteins. In the endoplasmic reticulum (ER), secretory proteins are packaged into COPII vesicles that largely exclude ER residents and misfolded proteins by mechanisms that remain unresolved. Here we combined biochemistry and genetics with correlative light and electron microscopy (CLEM) to explore how selectivity is achieved. Our data suggest that vesicle occupancy contributes to ER retention: in the absence of abundant cargo, nonspecific bulk flow increases. We demonstrate that ER leakage is influenced by vesicle size and cargo occupancy: overexpressing an inert cargo protein or reducing vesicle size restores sorting stringency. We propose that cargo recruitment into vesicles creates a crowded lumen that drives selectivity. Retention of ER residents thus derives in part from the biophysical process of cargo enrichment into a constrained spherical membrane-bound carrier.

In eukaryotic cells, proteins enter the secretory pathway at the endoplasmic reticulum (ER) as linear polypeptides and fold after translocation across or insertion into the membrane. If correct folding fails, many proteins are O-mannosylated inside the ER by an O-mannosyltransferase, the Pmt1p–Pmt2p complex. The consequences of this modification are controversial and the cellular role of the Pmt1p–Pmt2p complex in this respect is unclear. Here, we have identified the binding partners of yeast Pmt1p and Pmt2p. These include ER chaperones involved in oxidative protein folding; the Hrd1p complex, which is involved in ER-associated protein degradation (ERAD); and the p24 protein complex involved in ER export. The results suggest that the Pmt1p–Pmt2p complex participates in these processes. We tested this assumption in a functional assay and found that whereas the Pmt1p–Pmt2p complex promotes fast ER export of the GPI-anchored protein Gas1p, it retains the misfolded version Gas1*p and targets it to the Hrd1p complex for subsequent degradation. Our results reveal previously unknown cellular roles of the Pmt1p–Pmt2p complex in connection with the ERAD machinery and show its participation in ER protein quality control.

CLEM in yeast cells was used to characterize the formation of COPII vesicles. Their analysis suggests that the COPII inner coat defines the radius of curvature, whereas the outer coat facilitates vesicle fission. Coat organization and membrane biophysical properties determine the ultrastructure of vesicles and protein transport efficiency.

Crosslinked hyaluronic acid dermal fillers are widely used for non-permanent aesthetic enhancement, offering safe and effective solutions for facial volume restoration. Specific formulations are designed for targeted facial regions, with highly crosslinked hydrogels often recommended for volumizing areas such as the jawline, chin, and cheeks due to their structural properties. While elasticity and viscosity are commonly evaluated, broader comparative analyses remain limited. In this study, we assess five commercially available HA-based fillers with similar HA concentrations, all optimised for volume enhancement. Alongside widely used reference products, we evaluate BtHCROSS 2%^®, a novel formulation not previously compared to established fillers. We examined the degree of chemical modification, mechanical viscoelastic behaviour, susceptibility to enzymatic degradation by hyaluronidase, and injection force. While all tested fillers are suitable for volume restoration, BtHCROSS 2%^® demonstrates a distinctive combination of mechanical adaptability, structural support, enzymatic resistance, and low injection force, making it a versatile option for practitioners.