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Retapamulin-assisted ribosome profiling reveals the alternative bacterial proteome.
Mol. Cell 74, 481-493.e6 (2019)
Verlagsversion
Preprint
Forschungsdaten
DOI
PMC
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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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
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
Zeitschrift
Molecular Cell
Quellenangaben
Band: 74,
Heft: 3,
Seiten: 481-493.e6
Verlag
Elsevier
Verlagsort
50 Hampshire St, Floor 5, Cambridge, Ma 02139 Usa
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed
Institut(e)
Institute for Pancreatic Beta Cell Research (IPI)