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PreprintCited by 38RNA and protein synthesis mechanismsCellular transport and secretionEndoplasmic Reticulum Stress and Disease

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Structural insights into TRAP association with ribosome-Sec61 complex, and translocon inhibition by a CADA derivative

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Pauwels, Eva, Shewakramani, Neesha R., De Wijngaert, Brent, Camps, Anita, Provinciael, Becky, Stroobants, Joren, Kalies, Kai-Uwe, Hartmann, Enno, Maes, Piet, Vermeire, Kurt, Das, Kalyan·2022·Science Advances
DOI 10.1101/2022.09.28.509949PMC PMC9984176

Preprint — published as 10.1126/sciadv.adf0797

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Abstract

During co-translational translocation, the signal peptide of a nascent chain binds Sec61 translocon to initiate protein transport through the ER membrane. Our cryo-EM structure of ribosome-Sec61 shows binding of an ordered heterotetrameric TRranslocon-Associated Protein (TRAP) complex, in which TRAP-γ is anchored at two adjacent positions of 28S rRNA and interacts with ribosomal protein L38 and Sec61α/γ. Four transmembrane helices (TMHs) of TRAP-γ cluster with one C-terminal helix of each α, β, and δ subunits. The seven TMH bundle helps position a crescent-shaped trimeric TRAP–α/β/δ core in the ER lumen, facing the Sec61 channel. Further, our in vitro assay establishes the CADA derivative CK147 as a translocon inhibitor. A structure of ribosome-Sec61-CK147 reveals CK147 binding the channel and interacting with the plug helix from the lumenal side. The CK147-resistance mutations surround the inhibitor. These structures help in understanding the TRAP functions and provide a new Sec61 site for designing translocon inhibitors. Short Summary: Cryo-EM structures reveal TRAP binding to ribosome-Sec61 complex, and CK147 inhibiting Sec61 by arresting the plug helix inside the channel.

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Metadata

Authors
Pauwels, E., Shewakramani, N. R., De Wijngaert, B., Camps, A., Provinciael, B., Stroobants, J., Kalies, K.-U., Hartmann, E., Maes, P., Vermeire, K., & Das, K.
Affiliations
  • Rega Institute for Medical Research (BE)
  • University of Lübeck (DE)
Corresponding author
  • Vermeire, Kurt
  • Das, Kalyan
ORCID
Venue
Science Advances
Publisher
openRxiv
Published
2022-09-28
Volume
9
Issue
9
Pages
eadf0797-eadf0797
ISSN
2375-2548 (ISSN-L)
DOI
10.1101/2022.09.28.509949
PMC
PMC9984176
OpenAlex
W4323031742
Cited by
38
References
1
Citations by year
2026: 5 · 2025: 14 · 2024: 10 · 2023: 8 · 2022: 1
FWCI
3.30
Type
journal article
Language
en
Open access
Licence
cc-by-nc
Locations
  • Science Advances (publishedVersion, cc-by-nc)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
  • Lirias (KU Leuven) (submittedVersion, cc-by-nc-nd)
  • Lirias (KU Leuven) (submittedVersion, cc-by-nc)
Funders
KU Leuven
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Endoplasmic Reticulum Stress and Disease
Keywords
TRAP complex, Sec61 translocon, cotranslational translocation, signal peptide, 28S rRNA, transmembrane helices, endoplasmic reticulum membrane, protein trafficking
MeSH terms
SEC Translocation Channels, Calcium-Binding Proteins, Membrane Glycoproteins, Ribosomes, Receptors, Peptide, Receptors, Cytoplasmic and Nuclear, Cryoelectron Microscopy

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