Evidence for a cytoplasmic pool of ribosome-free mRNAs encoding inner membrane proteins in Escherichia coli

PLoS One. 2017 Aug 25;12(8):e0183862. doi: 10.1371/journal.pone.0183862. eCollection 2017.

Abstract

Translation-independent mRNA localization represents an emerging concept in cell biology. In Escherichia coli, mRNAs encoding integral membrane proteins (MPRs) are targeted to the membrane where they are translated by membrane associated ribosomes and the produced proteins are inserted into the membrane co-translationally. In order to better understand aspects of the biogenesis and localization of MPRs, we investigated their subcellular distribution using cell fractionation, RNA-seq and qPCR. The results show that MPRs are overrepresented in the membrane fraction, as expected, and depletion of the signal recognition particle-receptor, FtsY reduced the amounts of all mRNAs on the membrane. Surprisingly, however, MPRs were also found relatively abundant in the soluble ribosome-free fraction and their amount in this fraction is increased upon overexpression of CspE, which was recently shown to interact with MPRs. CspE also conferred a positive effect on the membrane-expression of integral membrane proteins. We discuss the possibility that the effects of CspE overexpression may link the intriguing subcellular localization of MPRs to the cytosolic ribosome-free fraction with their translation into membrane proteins and that the ribosome-free pool of MPRs may represent a stage during their targeting to the membrane, which precedes translation.

MeSH terms

  • Cytoplasm / metabolism*
  • Escherichia coli Proteins / genetics*
  • Membrane Proteins / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Ribosomes*
  • Sequence Analysis, RNA

Substances

  • Escherichia coli Proteins
  • Membrane Proteins
  • RNA, Messenger

Grants and funding

This work was supported by the Israel Science Foundation research grant (541/15), the Minerva Foundation with funding from the Federal German Ministry for Education and Research and by the Pasteur-Weizmann center to EB.