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Meydan, S.* ; Marks, J.* ; Klepacki, D.* ; Sharma, V. ; Baranov, P.V.* ; Firth, A.E.* ; Margus, T.* ; Kefi, A.* ; Vázquez-Laslop, N.* ; Mankin, A.S.*

Retapamulin-assisted ribosome profiling reveals the alternative bacterial proteome.

Mol. Cell 74, 481-493.e6 (2019)
Publ. Version/Full Text Preprint Research data DOI
Open Access Green as soon as Postprint is submitted to ZB.
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.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Alternative Initiation ; Arcb ; Internal Genes ; Retapamulin ; Ribosome Profiling ; Rpn ; Spea ; Translation Initiation; Biosynthetic Arginine Decarboxylase; Translational Initiation Sites; Escherichia-coli; Wide Analysis; Start Sites; In-vivo; Gene; Identification; Pleuromutilin; Proteins
ISSN (print) / ISBN 1097-2765
e-ISSN 1097-4164
Journal Molecular Cell
Quellenangaben Volume: 74, Issue: 3, Pages: 481-493.e6 Article Number: , Supplement: ,
Publisher Elsevier
Publishing Place 50 Hampshire St, Floor 5, Cambridge, Ma 02139 Usa
Reviewing status Peer reviewed
Institute(s) Institute for Pancreatic Beta Cell Research (IPI)