Retapamulin-Assisted Ribosome Profiling Reveals the Alternative Bacterial Proteome

Sezen Meydan, James Marks, Dorota Klepacki, Virag Sharma, Pavel V. Baranov, Andrew E. Firth, Tōnu Margus, Amira Kefi, Nora Vázquez-Laslop, Alexander S. Mankin

Research output: Contribution to journalArticlepeer-review

Abstract

The use of alternative translation initiation sites enables production of more than one protein from a single gene, thereby expanding the cellular proteome. Although several such examples have been serendipitously found in bacteria, genome-wide mapping of alternative translation start sites has been unattainable. We found that the antibiotic retapamulin specifically arrests initiating ribosomes at start codons of the genes. Retapamulin-enhanced Ribo-seq analysis (Ribo-RET)not only allowed mapping of conventional initiation sites at the beginning of the genes, but strikingly, it also revealed putative internal start sites in a number of Escherichia coli genes. Experiments demonstrated that the internal start codons can be recognized by the ribosomes and direct translation initiation in vitro and in vivo. Proteins, whose synthesis is initiated at internal in-frame and out-of-frame start sites, can be functionally important and contribute to the “alternative” bacterial proteome. The internal start sites may also play regulatory roles in gene expression.

Original languageEnglish
Pages (from-to)481-493.e6
JournalMolecular Cell
Volume74
Issue number3
DOIs
Publication statusPublished - 2 May 2019
Externally publishedYes

Keywords

  • alternative initiation
  • arcB
  • internal genes
  • retapamulin
  • ribosome profiling
  • rpn
  • speA
  • translation initiation

Fingerprint

Dive into the research topics of 'Retapamulin-Assisted Ribosome Profiling Reveals the Alternative Bacterial Proteome'. Together they form a unique fingerprint.

Cite this