Posttranslational insertion of small membrane proteins by the bacterial signal recognition particle

PLoS Biol. 2020 Sep 30;18(9):e3000874. doi: 10.1371/journal.pbio.3000874. eCollection 2020 Sep.

Abstract

Small membrane proteins represent a largely unexplored yet abundant class of proteins in pro- and eukaryotes. They essentially consist of a single transmembrane domain and are associated with stress response mechanisms in bacteria. How these proteins are inserted into the bacterial membrane is unknown. Our study revealed that in Escherichia coli, the 27-amino-acid-long model protein YohP is recognized by the signal recognition particle (SRP), as indicated by in vivo and in vitro site-directed cross-linking. Cross-links to SRP were also observed for a second small membrane protein, the 33-amino-acid-long YkgR. However, in contrast to the canonical cotranslational recognition by SRP, SRP was found to bind to YohP posttranslationally. In vitro protein transport assays in the presence of a SecY inhibitor and proteoliposome studies demonstrated that SRP and its receptor FtsY are essential for the posttranslational membrane insertion of YohP by either the SecYEG translocon or by the YidC insertase. Furthermore, our data showed that the yohP mRNA localized preferentially and translation-independently to the bacterial membrane in vivo. In summary, our data revealed that YohP engages an unique SRP-dependent posttranslational insertion pathway that is likely preceded by an mRNA targeting step. This further highlights the enormous plasticity of bacterial protein transport machineries.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Escherichia coli / metabolism
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Membrane Proteins / metabolism*
  • Models, Biological
  • Protein Binding
  • Protein Biosynthesis
  • Protein Processing, Post-Translational*
  • Protein Transport
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • SEC Translocation Channels / metabolism
  • Signal Recognition Particle / metabolism*

Substances

  • Escherichia coli Proteins
  • Membrane Proteins
  • RNA, Messenger
  • SEC Translocation Channels
  • Signal Recognition Particle