SRP, FtsY, DnaK and YidC Are Required for the Biogenesis of the E. coli Tail-Anchored Membrane Proteins DjlC and Flk

Markus Peschke, Mélanie Le Goff, Gregory M. Koningstein, Alexandros Karyolaimos, Jan Willem de Gier, Peter van Ulsen, Joen Luirink*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Tail-anchored membrane proteins (TAMPs) are relatively simple membrane proteins characterized by a single transmembrane domain (TMD) at their C-terminus. Consequently, the hydrophobic TMD, which acts as a subcellular targeting signal, emerges from the ribosome only after termination of translation precluding canonical co-translational targeting and membrane insertion. In contrast to the well-studied eukaryotic TAMPs, surprisingly little is known about the cellular components that facilitate the biogenesis of bacterial TAMPs. In this study, we identify DjlC and Flk as bona fide Escherichia coli TAMPs and show that their TMDs are necessary and sufficient for authentic membrane targeting of the fluorescent reporter mNeonGreen. Using strains conditional for the expression of known E. coli membrane targeting and insertion factors, we demonstrate that the signal recognition particle (SRP), its receptor FtsY, the chaperone DnaK and insertase YidC are each required for efficient membrane localization of both TAMPs. A close association between the TMD of DjlC and Flk with both the Ffh subunit of SRP and YidC was confirmed by site-directed in vivo photo-crosslinking. In addition, our data suggest that the hydrophobicity of the TMD correlates with the dependency on SRP for efficient targeting.

Original languageEnglish
Pages (from-to)389-403
Number of pages15
JournalJournal of Molecular Biology
Volume430
Issue number3
DOIs
Publication statusAccepted/In press - 1 Jan 2017

Keywords

  • E. coli
  • Membrane insertion
  • Membrane protein
  • Membrane targeting
  • Tail-anchored

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