Antibiotic resistance by high-level intrinsic suppression of a frameshift mutation in an essential gene
Copyright © 2020 the Author(s). Published by PNAS..
A fundamental feature of life is that ribosomes read the genetic code in messenger RNA (mRNA) as triplets of nucleotides in a single reading frame. Mutations that shift the reading frame generally cause gene inactivation and in essential genes cause loss of viability. Here we report and characterize a +1-nt frameshift mutation, centrally located in rpoB, an essential gene encoding the beta-subunit of RNA polymerase. Mutant Escherichia coli carrying this mutation are viable and highly resistant to rifampicin. Genetic and proteomic experiments reveal a very high rate (5%) of spontaneous frameshift suppression occurring on a heptanucleotide sequence downstream of the mutation. Production of active protein is stimulated to 61-71% of wild-type level by a feedback mechanism increasing translation initiation. The phenomenon described here could have broad significance for predictions of phenotype from genotype. Several frameshift mutations have been reported in rpoB in rifampicin-resistant clinical isolates of Mycobacterium tuberculosis (Mtb). These mutations have never been experimentally validated, and no mechanisms of action have been proposed. This work shows that frameshift mutations in rpoB can be a mutational mechanism generating antibiotic resistance. Our analysis further suggests that genetic elements supporting productive frameshifting could rapidly evolve de novo, even in essential genes.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2020 |
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| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:117 |
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| Enthalten in: |
Proceedings of the National Academy of Sciences of the United States of America - 117(2020), 6 vom: 11. Feb., Seite 3185-3191 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Huseby, Douglas L [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 11.05.2020 Date Revised 11.05.2020 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1073/pnas.1919390117 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM305887181 |
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| 245 | 1 | 0 | |a Antibiotic resistance by high-level intrinsic suppression of a frameshift mutation in an essential gene |
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| 500 | |a Date Revised 11.05.2020 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2020 the Author(s). Published by PNAS. | ||
| 520 | |a A fundamental feature of life is that ribosomes read the genetic code in messenger RNA (mRNA) as triplets of nucleotides in a single reading frame. Mutations that shift the reading frame generally cause gene inactivation and in essential genes cause loss of viability. Here we report and characterize a +1-nt frameshift mutation, centrally located in rpoB, an essential gene encoding the beta-subunit of RNA polymerase. Mutant Escherichia coli carrying this mutation are viable and highly resistant to rifampicin. Genetic and proteomic experiments reveal a very high rate (5%) of spontaneous frameshift suppression occurring on a heptanucleotide sequence downstream of the mutation. Production of active protein is stimulated to 61-71% of wild-type level by a feedback mechanism increasing translation initiation. The phenomenon described here could have broad significance for predictions of phenotype from genotype. Several frameshift mutations have been reported in rpoB in rifampicin-resistant clinical isolates of Mycobacterium tuberculosis (Mtb). These mutations have never been experimentally validated, and no mechanisms of action have been proposed. This work shows that frameshift mutations in rpoB can be a mutational mechanism generating antibiotic resistance. Our analysis further suggests that genetic elements supporting productive frameshifting could rapidly evolve de novo, even in essential genes | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a antibiotic resistance | |
| 650 | 4 | |a evolution | |
| 650 | 4 | |a frameshift suppression | |
| 650 | 4 | |a gene regulation | |
| 650 | 4 | |a rpoB | |
| 650 | 7 | |a Escherichia coli Proteins |2 NLM | |
| 650 | 7 | |a rpoB protein, E coli |2 NLM | |
| 650 | 7 | |a DNA-Directed RNA Polymerases |2 NLM | |
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| 700 | 1 | |a Brandis, Gerrit |e verfasserin |4 aut | |
| 700 | 1 | |a Praski Alzrigat, Lisa |e verfasserin |4 aut | |
| 700 | 1 | |a Hughes, Diarmaid |e verfasserin |4 aut | |
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