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Volume 170, Issue 2

Identification of novel tail-anchored membrane proteins integrated by the bacterial twin-arginine translocase Open Access

Abstract

The twin-arginine protein transport (Tat) system exports folded proteins across the cytoplasmic membranes of prokaryotes and the energy transducing-membranes of plant thylakoids and mitochondria. Proteins are targeted to the Tat machinery by N-terminal signal peptides with a conserved twin-arginine motif, and some substrates are exported as heterodimers where the signal peptide is present on one of the partner proteins. A subset of Tat substrates is found in the membrane. Tat-dependent membrane proteins usually have large globular domains and a single transmembrane helix present at the N- or C-terminus. Five Tat substrates that have C-terminal transmembrane helices have previously been characterized in the model bacterium . Each of these is an iron–sulfur cluster-containing protein involved in electron transfer from hydrogen or formate. Here we have undertaken a bioinformatic search to identify further tail-anchored Tat substrates encoded in bacterial genomes. Our analysis has revealed additional tail-anchored iron–sulfur proteins associated in modules with either a -type cytochrome or a quinol oxidase. We also identified further candidate tail-anchored Tat substrates, particularly among members of the actinobacterial phylum, that are not predicted to contain cofactors. Using reporter assays, we show experimentally that six of these have both N-terminal Tat signal peptides and C-terminal transmembrane helices. The newly identified proteins include a carboxypeptidase and a predicted protease, and four sortase substrates for which membrane integration is a prerequisite for covalent attachment to the cell wall.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): bioinformatics , membrane protein , tail-anchored , Tat pathway and twin-arginine signal peptide
Funding
This study was supported by the:
  • Medical Research Council (Award MR/S009213/1)
    • Principle Award Recipient: TracyPalmer
  • Biotechnology and Biological Sciences Research Council (Award BB/S005307/1)
    • Principle Award Recipient: TracyPalmer
© 2024 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2024-02-16
2024-05-20
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Identification of novel tail-anchored membrane proteins integrated by the bacterial twin-arginine translocase
Microbiology 170, 001431 (2024); https://doi.org/10.1099/mic.0.001431
/content/journal/micro/10.1099/mic.0.001431
/content/journal/micro/10.1099/mic.0.001431
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