title: Mammalian OS-9 is upregulated in response to endoplasmic reticulum stress and facilitates ubiquitination of misfolded glycoproteins.
abstract: Proteins that fail to fold or assemble with partner subunits are selectively removed from the endoplasmic reticulum (ER) via the ER-associated degradation (ERAD) pathway. Proteins selected for ERAD are polyubiquitinated and retrotranslocated into the cytosol for degradation by the proteasome. Although it is unclear how proteins are initially identified by the ERAD system in mammalian cells, OS-9 was recently proposed to play a key role in this process. Here we show that OS-9 is upregulated in response to ER stress and is associated both with components of the ERAD machinery and with ERAD substrates. Using RNA interference, we show that OS-9 is required for efficient ubquitination of glycosylated ERAD substrates, suggesting that it helps transfer misfolded proteins to the ubiquitination machinery. We also find that OS-9 binds to a misfolded nonglycosylated protein destined for ERAD, but not to the properly folded wild-type protein. Surprisingly, however, OS-9 is not required for ubiquitination or degradation of this nonglycosylated ERAD substrate. We propose a model in which OS-9 recognises terminally misfolded proteins via polypeptide-based rather than glycan-based signals, but is only required for transferring those bearing N-glycans to the ubiquitination machinery.
authors.affiliation: Faculty of Life Sciences, University of Manchester, The Michael Smith Building, Oxford Road, Manchester M13 9PT, UK.
authors.forename: Felicity
authors.initials: F
authors.lastname: Alcock
authors.forename: Eileithyia
authors.initials: E
authors.lastname: Swanton
mesh_terms: D000818:Animals; D019295:Computational Biology; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000473:pathology*; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D037102:Lectins; D008322:Mammals / Q000378:metabolism*; D008954:Models, Biological; D009186:Myelin Sheath / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism*; D025821:Polyubiquitin / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D011489:Protein Denaturation; D017510:Protein Folding / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013312:Stress, Physiological; D019284:Thapsigargin / Q000494:pharmacology; D014162:Transfection; D054875:Ubiquitination / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D037102:Lectins; D009363:Neoplasm Proteins; C098898:OS9 protein, human; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D000515:alpha 1-Antitrypsin; D025821:Polyubiquitin; D019284:Thapsigargin