PXD035475-1

PXD035475 is an original dataset announced via ProteomeXchange.

Title	Molecular snapshots of translation and protein translocation at the ER membrane
Description	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. While individual components of this machine have been studied at high resolution in isolation, insights into their interplay in the native membrane remain limited. Here, we use electron cryo-tomography (cryo-ET), extensive classification and molecular modeling to capture molecular resolution snapshots of mRNA translation and protein maturation at the ER membrane. Comprehensively recapitulating the translational elongation cycle, we identify a highly abundant classical pre-translocation (PRE) intermediate with eEF1a in an extended conformation following the decoding state, which indicates that eEF1a remains ribosome-associated after GTP-hydrolysis during proofreading. The complete atomic structure of the most abundant ER translocon variant comprising the protein-conducting channel Sec61, the translocon-associated protein complex (TRAP) and the oligosaccharyltransferase complex A (OSTA) reveals the molecular framework for signal peptide (SP) membrane insertion and protein N-glycosylation. Associated with OSTA we observe stoichiometric and sub-stoichiometric cofactors, likely including protein isomerases. Collectively, comprehensive analysis of ER-associated protein biogenesis ex vivo reveals numerous mechanistic insights advancing beyond the study of isolated components.
HostingRepository	PRIDE
AnnounceDate	2023-02-03
AnnouncementXML	Submission_2023-02-03_07:26:09.125.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	RichardScheltema
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2022-07-21 03:39:19	ID requested
⏵ 1	2023-02-03 07:26:09	announced
2	2023-11-14 07:38:34	announced	2023-11-14: Updated project metadata.

Gemmer M, Chaillet ML, van Loenhout J, Cuevas Arenas R, Vismpas D, Gr, ö, llers-Mulderij M, Koh FA, Albanese P, Scheltema RA, Howes SC, Kotecha A, Fedry J, F, ö, rster F, Visualization of translation and protein biogenesis at the ER membrane. Nature, 614(7946):160-167(2023) [pubmed]

submitter keyword: Ribosome, ER membrane, elongation factor 1

RichardScheltema
contact affiliation	Utrecht University
contact email	r.a.scheltema@uu.nl
lab head
RichardScheltema
contact affiliation	Utrecht University
contact email	r.a.scheltema@uu.nl
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/02/PXD035475

PRIDE project URI

• PRIDE
  1. PXD035475
     1. Label: PRIDE project
     2. Name: Molecular snapshots of translation and protein translocation at the ER membrane