Fine interaction profiling of VemP and mechanisms responsible for its translocation-coupled arrest-cancelation
- Kyoto University, Japan
Abstract
Bacterial cells utilize monitoring substrates, which undergo force-sensitive translation elongation arrest, to feedback-regulate a Sec-related gene. Vibrio alginolyticus VemP controls the expression of SecD/F that stimulates a late step of translocation by undergoing export-regulated elongation arrest. Here, we attempted at delineating the pathway of the VemP nascent-chain interaction with Sec-related factors, and identified the signal recognition particle (SRP) and PpiD (a membrane-anchored periplasmic chaperone) in addition to other translocon components and a ribosomal protein as interacting partners. Our results showed that SRP is required for the membrane-targeting of VemP, whereas PpiD acts cooperatively with SecD/F in the translocation and arrest-cancelation of VemP. We also identified the conserved Arg-85 residue of VemP as a crucial element that confers PpiD-dependence to VemP and plays an essential role in the regulated arrest-cancelation. We propose a scheme of the arrest-cancelation processes of VemP, which likely monitors late steps in the protein translocation pathway.
Data availability
All data generated and analyzed during this study are included in the manuscript and supporting files. Source data files have been provided for Figures 2, 3 and 4, Figure 2-figure supplements 1, 3 and 4 and Figure 5-figure supplements 1 and 2.
Article and author information
Author details
Funding
Japan Society for the Promotion of Science (18H06047)
- Ryoji Miyazaki
Japan Society for the Promotion of Science (19K21179)
- Ryoji Miyazaki
Japan Society for the Promotion of Science (20K15715)
- Ryoji Miyazaki
Japan Society for the Promotion of Science (15H01532)
- Yoshinori Akiyama
Japan Society for the Promotion of Science (18H02404)
- Yoshinori Akiyama
Japan Society for the Promotion of Science (17H05666)
- Hiroyuki Mori
Japan Society for the Promotion of Science (17H05879)
- Hiroyuki Mori
Japan Society for the Promotion of Science (17K07334)
- Hiroyuki Mori
Japan Society for the Promotion of Science (20K06556)
- Hiroyuki Mori
The funders had no role in study design, data collection and interpretation.
Copyright
© 2020, Miyazaki et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Fine interaction profiling of VemP and mechanisms responsible for its translocation-coupled arrest-cancelation
eLife 9:e62623.
https://doi.org/10.7554/eLife.62623
