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

We follow the cotranslational biosynthesis of three multispanning 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 m...

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Bibliographic Details
Published in:eLife 2021-02, Vol.10
Main Authors: Nicolaus, Felix, Metola, Ane, Mermans, Daphne, Liljenström, Amanda, Krč, Ajda, Abdullahi, Salmo Mohammed, Zimmer, Matthew, Miller Iii, Thomas F, von Heijne, Gunnar
Format: Article
Language:English
Subjects:
Amino Acid Motifs
Analysis
Arrests
Biochemistry and Chemical Biology
biogenesis
BtuC
Cell Membrane - chemistry
Cell Membrane - genetics
Cell Membrane - metabolism
cotranslational
EmrE
Escherichia coli
Escherichia coli - chemistry
Escherichia coli - genetics
Escherichia coli - metabolism
Escherichia coli Proteins - chemistry
Escherichia coli Proteins - genetics
Escherichia coli Proteins - metabolism
Gene mutations
GlpG
Hydrophobicity
Integration
Lipid bilayers
Membrane lipids
membrane protein
Membrane proteins
Mutation
Physiological aspects
Polypeptides
Protein Biosynthesis
Protein Domains
Proteins
Structural Biology and Molecular Biophysics
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Summary:We follow the cotranslational biosynthesis of three multispanning 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.
ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.64302