9OG0

Cryo-EM structure of OS9-SEL1L-HRD1 dimer

Summary for 9OG0

• Entry DOI: 10.2210/pdb9og0/pdb
• EMDB information: 70448
• Descriptor: E3 ubiquitin-protein ligase synoviolin, Protein sel-1 homolog 1, Isoform 2 of Protein OS-9 (3 entities in total)
• Functional Keywords: erad, sel1l, os9, hrd1, membrane protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 6
• Total formula weight: 451030.17

Authors

Lin, L.,Maldosevic, E.,Jomaa, A.,Qi, L. (deposition date: 2025-04-30, release date: 2025-07-30)

Primary citation

Lin, L.L.,Maldosevic, E.,Zhou, L.E.,Jomaa, A.,Qi, L.
Structural basis and pathological implications of the dimeric OS9-SEL1L-HRD1 ERAD Core Complex.
Biorxiv, 2025

PubMed Abstract: The SEL1L-HRD1 complex represents the most conserved branch of endoplasmic reticulum (ER)-associated degradation (ERAD), a critical pathway that clears misfolded proteins to maintain ER proteostasis. However, the molecular organization and pathogenic mechanisms of mammalian ERAD have remained elusive. Here, we report the first cryo-EM structure of the core mammalian ERAD complex, comprising the ER lectin OS9, SEL1L, and the E3 ubiquitin ligase HRD1. The structure, validated by mutagenesis and crosslinking assays, reveals a dimeric assembly of the core complex in which SEL1L and OS9 form a claw-like configuration in the ER lumen that mediates substrate engagement, while HRD1 dimerizes within the membrane to facilitate substrate translocation. Pathogenic SEL1L mutations at the SEL1L-OS9 (Gly585Asp) and SEL1L-HRD1 (Ser658Pro) interfaces disrupt complex formation and impair ERAD activity. A newly identified disease-associated HRD1 variant (Ala91Asp), located in transmembrane helix 3, impairs HRD1 dimerization and substrate processing, underscoring the functional necessity of this interface and HRD1 dimerization. Finally, the structure also reveals two methionine-rich crevices flanking the HRD1 dimer, suggestive of substrate-conducting channels analogous to those in the ER membrane protein complex (EMC). These findings establish a structural framework for mammalian ERAD and elucidate how mutations destabilizing this machinery contribute to human disease.

PubMed: 40661598
DOI: 10.1101/2025.06.13.659592

Experimental method

ELECTRON MICROSCOPY (3.64 Å)