8B6Z

CryoEM Structure of Extended eEF1A bound to the Ribosome in the Classical Pre State

Summary for 8B6Z

• Entry DOI: 10.2210/pdb8b6z/pdb
• EMDB information: 15893
• Descriptor: Elongation factor 1-alpha 2, 28S ribosomal RNA (2 entities in total)
• Functional Keywords: translation, protein biogenesis, elongation factor, ribosome
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 2
• Total formula weight: 1690767.23

Authors

Gemmer, M.,Fedry, J.M.M.,Forster, F.G. (deposition date: 2022-09-27, release date: 2022-12-07, Last modification date: 2024-07-24)

Primary citation

Gemmer, M.,Chaillet, M.L.,van Loenhout, J.,Cuevas Arenas, R.,Vismpas, D.,Grollers-Mulderij, M.,Koh, F.A.,Albanese, P.,Scheltema, R.A.,Howes, S.C.,Kotecha, A.,Fedry, J.,Forster, F.
Visualization of translation and protein biogenesis at the ER membrane.
Nature, 614:160-167, 2023

PubMed Abstract: The dynamic ribosome-translocon complex, which resides at the endoplasmic reticulum (ER) membrane, produces a major fraction of the human proteome. It governs the synthesis, translocation, membrane insertion, N-glycosylation, folding and disulfide-bond formation of nascent proteins. Although individual components of this machinery have been studied at high resolution in isolation, insights into their interplay in the native membrane remain limited. Here we use cryo-electron tomography, extensive classification and molecular modelling to capture snapshots of mRNA translation and protein maturation at the ER membrane at molecular resolution. We identify a highly abundant classical pre-translocation intermediate with eukaryotic elongation factor 1a (eEF1a) in an extended conformation, suggesting that eEF1a may remain associated with the ribosome after GTP hydrolysis during proofreading. At the ER membrane, distinct polysomes bind to different ER translocons specialized in the synthesis of proteins with signal peptides or multipass transmembrane proteins with the translocon-associated protein complex (TRAP) present in both. The near-complete atomic model of the most abundant ER translocon variant comprising the protein-conducting channel SEC61, TRAP and the oligosaccharyltransferase complex A (OSTA) reveals specific interactions of TRAP with other translocon components. We observe stoichiometric and sub-stoichiometric cofactors associated with OSTA, which are likely to include protein isomerases. In sum, we visualize ER-bound polysomes with their coordinated downstream machinery.

PubMed: 36697828
DOI: 10.1038/s41586-022-05638-5

Experimental method

ELECTRON MICROSCOPY (2.9 Å)