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Transient erythromycin resistance phenotype associated with peptidyl-tRNA drop-off on early UGG and GGG codons

Macvanin, Mirjana ; Gonzalez de Valdivia, Ernesto I LU orcid ; Ardell, David H and Isaksson, Leif A (2007) In Journal of Bacteriology 189(24). p.8993-9000
Abstract

Expression of minigenes encoding tetra- or pentapeptides MXLX or MXLXV (E peptides), where X is a nonpolar amino acid, renders cells erythromycin resistant whereas expression of minigenes encoding tripeptide MXL does not. By using a 3A' reporter gene system beginning with an E-peptide-encoding sequence, we asked whether the codons UGG and GGG, which are known to promote peptidyl-tRNA drop-off at early positions in mRNA, would result in a phenotype of erythromycin resistance if located after this sequence. We find that UGG or GGG, at either position +4 or +5, without a following stop codon, is associated with an erythromycin resistance phenotype upon gene induction. Our results suggest that, while a stop codon at +4 gives a tripeptide... (More)

Expression of minigenes encoding tetra- or pentapeptides MXLX or MXLXV (E peptides), where X is a nonpolar amino acid, renders cells erythromycin resistant whereas expression of minigenes encoding tripeptide MXL does not. By using a 3A' reporter gene system beginning with an E-peptide-encoding sequence, we asked whether the codons UGG and GGG, which are known to promote peptidyl-tRNA drop-off at early positions in mRNA, would result in a phenotype of erythromycin resistance if located after this sequence. We find that UGG or GGG, at either position +4 or +5, without a following stop codon, is associated with an erythromycin resistance phenotype upon gene induction. Our results suggest that, while a stop codon at +4 gives a tripeptide product (MIL) and erythromycin sensitivity, UGG or GGG codons at the same position give a tetrapeptide product (MILW or MILG) and phenotype of erythromycin resistance. Thus, the drop-off event on GGG or UGG codons occurs after incorporation of the corresponding amino acid into the growing peptide chain. Drop-off gives rise to a peptidyl-tRNA where the peptide moiety functionally mimics a minigene peptide product of the type previously associated with erythromycin resistance. Several genes in Escherichia coli fulfill the requirements of high mRNA expression and an E-peptide sequence followed by UGG or GGG at position +4 or +5 and should potentially be able to give an erythromycin resistance phenotype.

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author
; ; and
publishing date
type
Contribution to journal
publication status
published
keywords
Anti-Bacterial Agents, Codon, Drug Resistance, Bacterial, Erythromycin, Escherichia coli, Genes, Reporter, Oligopeptides, Protein Biosynthesis, RNA, Transfer, Amino Acyl, Staphylococcal Protein A, Journal Article, Research Support, Non-U.S. Gov't
in
Journal of Bacteriology
volume
189
issue
24
pages
8 pages
publisher
American Society for Microbiology
external identifiers
  • pmid:17951392
  • scopus:37449027381
ISSN
0021-9193
DOI
10.1128/JB.01004-07
language
English
LU publication?
no
id
ccb2aac4-aaf2-48b3-bd27-e636facf535a
date added to LUP
2018-01-13 11:53:48
date last changed
2024-04-29 02:00:33
@article{ccb2aac4-aaf2-48b3-bd27-e636facf535a,
  abstract     = {{<p>Expression of minigenes encoding tetra- or pentapeptides MXLX or MXLXV (E peptides), where X is a nonpolar amino acid, renders cells erythromycin resistant whereas expression of minigenes encoding tripeptide MXL does not. By using a 3A' reporter gene system beginning with an E-peptide-encoding sequence, we asked whether the codons UGG and GGG, which are known to promote peptidyl-tRNA drop-off at early positions in mRNA, would result in a phenotype of erythromycin resistance if located after this sequence. We find that UGG or GGG, at either position +4 or +5, without a following stop codon, is associated with an erythromycin resistance phenotype upon gene induction. Our results suggest that, while a stop codon at +4 gives a tripeptide product (MIL) and erythromycin sensitivity, UGG or GGG codons at the same position give a tetrapeptide product (MILW or MILG) and phenotype of erythromycin resistance. Thus, the drop-off event on GGG or UGG codons occurs after incorporation of the corresponding amino acid into the growing peptide chain. Drop-off gives rise to a peptidyl-tRNA where the peptide moiety functionally mimics a minigene peptide product of the type previously associated with erythromycin resistance. Several genes in Escherichia coli fulfill the requirements of high mRNA expression and an E-peptide sequence followed by UGG or GGG at position +4 or +5 and should potentially be able to give an erythromycin resistance phenotype.</p>}},
  author       = {{Macvanin, Mirjana and Gonzalez de Valdivia, Ernesto I and Ardell, David H and Isaksson, Leif A}},
  issn         = {{0021-9193}},
  keywords     = {{Anti-Bacterial Agents; Codon; Drug Resistance, Bacterial; Erythromycin; Escherichia coli; Genes, Reporter; Oligopeptides; Protein Biosynthesis; RNA, Transfer, Amino Acyl; Staphylococcal Protein A; Journal Article; Research Support, Non-U.S. Gov't}},
  language     = {{eng}},
  number       = {{24}},
  pages        = {{8993--9000}},
  publisher    = {{American Society for Microbiology}},
  series       = {{Journal of Bacteriology}},
  title        = {{Transient erythromycin resistance phenotype associated with peptidyl-tRNA drop-off on early UGG and GGG codons}},
  url          = {{http://dx.doi.org/10.1128/JB.01004-07}},
  doi          = {{10.1128/JB.01004-07}},
  volume       = {{189}},
  year         = {{2007}},
}