Derlin rhomboid pseudoproteases employ substrate engagement and lipid distortion to enable the retrotranslocation of ERAD membrane substrates

Nearly one-third of proteins are initially targeted to the endoplasmic reticulum (ER) membrane, where they are correctly folded and then delivered to their final cellular destinations. To prevent the accumulation of misfolded membrane proteins, ER-associated degradation (ERAD) moves these clients from the ER membrane to the cytosol, a process known as retrotranslocation. Our recent work in Saccharomyces cerevisiae reveals a derlin rhomboid pseudoprotease, Dfm1, is involved in the retrotranslocation of ubiquitinated ERAD membrane substrates. In this study, we identify conserved residues of Dfm1 that are critical for retrotranslocation. We find several retrotranslocation-deficient Loop 1 mutants that display impaired binding to membrane substrates. Furthermore, Dfm1 possesses lipid thinning function to facilitate in the removal of ER membrane substrates, and this feature is conserved in its human homolog, Derlin-1, further implicating that derlin-mediated retrotranslocation is a well-conserved process. [Display omitted] •Dfm1 selectively binds ERAD-targeted membrane substrates•Polyubiquitin chains bind directly to Cdc48 recruited by Dfm1•Derlin lipid thinning facilitates removal of integral membrane substrates in the ER•Substrate engagement and lipid thinning are conserved derlin features ER-associated degradation is a conserved pathway of protein quality control that requires the retrotranslocation of ubiquitinated substrates from the ER to the cytoplasm for degradation. Nejatfard et al. show that derlin rhomboid pseudoproteases mediate the retrotranslocation of misfolded membrane substrates via a mechanism that is conserved from yeast to humans.