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Residue-by-residue analysis of cotranslational membrane protein integration in vivo

Nicolaus, Felix and Metola, Ane and Mermans, Daphne and Liljenström, Amanda and Krč, Ajda and Abdullahi, Salmo Mohammed and Zimmer, Matthew and Miller, Thomas F., III and Von Heijne, Gunnar (2021) Residue-by-residue analysis of cotranslational membrane protein integration in vivo. eLife, 10 . Art. No. e64302. ISSN 2050-084X.

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We follow the cotranslational biosynthesis of three multispanning Escherichia coli inner membrane proteins in vivo using high-resolution force profile analysis. The force profiles show that the nascent chain is subjected to rapidly varying pulling forces during translation and reveal unexpected complexities in the membrane integration process. We find that an N-terminal cytoplasmic domain can fold in the ribosome exit tunnel before membrane integration starts, that charged residues and membrane-interacting segments such as re-entrant loops and surface helices flanking a transmembrane helix (TMH) can advance or delay membrane integration, and that point mutations in an upstream TMH can affect the pulling forces generated by downstream TMHs in a highly position-dependent manner, suggestive of residue-specific interactions between TMHs during the integration process. Our results support the ‘sliding’ model of translocon-mediated membrane protein integration, in which hydrophobic segments are continually exposed to the lipid bilayer during their passage through the SecYEG translocon.

Item Type:Article
Related URLs:
URLURL TypeDescription Letter Response Paper
Nicolaus, Felix0000-0001-9230-8544
Metola, Ane0000-0002-2885-7634
Mermans, Daphne0000-0001-6001-5608
Zimmer, Matthew0000-0002-1437-2636
Miller, Thomas F., III0000-0002-1882-5380
Von Heijne, Gunnar0000-0002-4490-8569
Additional Information:© 2021 Nicolaus et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Received: 23 October 2020; Accepted: 05 February 2021; Published: 08 February 2021. We thank Dr. Rickard Hedman (Stockholm University) for programming and maintenance of the EasyQuant software. This work was supported by grants from the Knut and Alice Wallenberg Foundation (2017.0323), the Novo Nordisk Fund (NNF18OC0032828), and the Swedish Research Council (621-2014-3713) to GvH, from a Marie Curie Initial Training Network Grant (Horizon 2020, Protein-Factory 642863) to FN, and from NIGMS, National Institutes of Health, (R01GM125063) to TFM and MZ. This work used the Extreme Science and Engineering Discovery Environment (XSEDE) Bridges computer at PSC through allocation TG-MCB160013. XSEDE is supported by National Science Foundation grant number ACI-1548562. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Data availability: All fFL values measured in this study are included as figures Source Data. Author contributions: Felix Nicolaus, Ane Metola, Daphne Mermans, Formal analysis, Supervision, Investigation, Visualization, Methodology, Writing - review and editing; Amanda Liljenstroöm, Ajda Krč, Salmo Mohammed Abdullahi, Investigation; Matthew Zimmer, Formal analysis, Funding acquisition, Investigation, Writing - review and editing; Thomas F Miller III, Conceptualization, Funding acquisition, Writing - review and editing; Gunnar von Heijne, Conceptualization, Resources, Formal analysis, Supervision, Funding acquisition, Validation, Visualization, Methodology, Writing - original draft, Project administration, Writing - review and editing. The authors declare that no competing interests exist.
Funding AgencyGrant Number
Knut and Alice Wallenberg Foundation2017.0323
Novo Nordisk FundNNF18OC0032828
Swedish Research Council621-2014-3713
Marie Curie Fellowship642863
Record Number:CaltechAUTHORS:20200929-122616290
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Official Citation:Residue-by-residue analysis of cotranslational membrane protein integration in vivo eLife 2021; 10:e64302; DOI: 10.7554/eLife.64302
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:105636
Deposited By: Tony Diaz
Deposited On:29 Sep 2020 20:12
Last Modified:18 Feb 2021 17:35

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