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  Co‐translational insertion and topogenesis of bacterial membrane proteins monitored in real time

Mercier, E., Wintermeyer, W., & Rodnina, M. V. (2020). Co‐translational insertion and topogenesis of bacterial membrane proteins monitored in real time. EMBO Journal, e104054. doi:10.15252/embj.2019104054.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0006-8AB0-7 Version Permalink: http://hdl.handle.net/21.11116/0000-0006-8ABE-9
Genre: Journal Article

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Mercier, E.1, Author              
Wintermeyer, W.1, Author              
Rodnina, M. V.1, Author              
Affiliations:
1Department of Physical Biochemistry, MPI for Biophysical Chemistry, Max Planck Society, ou_578598              

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Free keywords: In vitro translation; positive-inside rule; real-time translation kinet- ics; ribosome; transmembrane segment inversion
 Abstract: Integral membrane proteins insert into the bacterial inner membrane co‐translationally via the translocon. Transmembrane (TM ) segments of nascent proteins adopt their native topological arrangement with the N‐terminus of the first TM (TM 1) oriented to the outside (type I) or the inside (type II ) of the cell. Here, we study TM 1 topogenesis during ongoing translation in a bacterial in vitro system, applying real‐time FRET and protease protection assays. We find that TM 1 of the type I protein LepB reaches the translocon immediately upon emerging from the ribosome. In contrast, the type II protein EmrD requires a longer nascent chain before TM 1 reaches the translocon and adopts its topology by looping inside the ribosomal peptide exit tunnel. Looping presumably is mediated by interactions between positive charges at the N‐terminus of TM 1 and negative charges in the tunnel wall. Early TM 1 inversion is abrogated by charge reversal at the N‐terminus. Kinetic analysis also shows that co‐translational membrane insertion of TM 1 is intrinsically rapid and rate‐limited by translation. Thus, the ribosome has an important role in membrane protein topogenesis.

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Language(s): eng - English
 Dates: 2020-04-20
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.15252/embj.2019104054
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Title: EMBO Journal
Source Genre: Journal
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Pages: 13 Volume / Issue: - Sequence Number: e104054 Start / End Page: - Identifier: -