Transient erythromycin resistance phenotype associated with peptidyl-tRNA drop-off on early UGG and GGG codons
(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
- Macvanin, Mirjana ; Gonzalez de Valdivia, Ernesto I LU ; Ardell, David H and Isaksson, Leif A
- publishing date
- 2007-12
- 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}}, }