The DOI matches its Crossref record.
Effects of mutagenesis of a conserved base-paired site near the decoding region of Escherichia coli 16 S ribosomal RNA
Abstract
Eleven of 15 possible single and double mutations were constructed in a cloned copy of Escherichia coli 16 S rDNA at a base-paired site, 1409-1491. Expression of any of these mutations was detrimental to the growth of E. coli. Mutations that substituted unpaired purine bases were lethal in the system described. Otherwise, the degree of detrimental effect on growth-rate was not directly correlated with specific rRNA primary or secondary structures. Using reverse transcription of rRNA isolated from subunits or 70 S ribosomes, we were able to determine the amount of mutant rRNA used in translation. From these experiments, we found that the lethal mutations appeared to be selectively excluded from the pool of 70 S ribosomes following expression from a repressible plasmid. In contrast, a non-lethal mutation was present in subunits, ribosomes and polysomes in approximately equal amounts. Mutations that disrupted base-pairing were found to confer varying levels of resistance to nine aminoglycosides, including four neomycins, two kanamycins, gentamicin, apramycin and hygromycin. A high frequency of reversion from resistant and slow-growth phenotypes due to a host mutation was observed.
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In the citing paper
“Neither of these residues is near the binding site of HygB, and the reported loss of binding of the antibiotic must be due to an allosteric effect caused by disruption of the G1491:C1409 base pair, as the authors themselves suggest (De Stasio and Dahlberg 1990”
From The Structural Basis for the Action of the Antibiotics Tetracycline, Pactamycin, and Hygromycin B on the 30S Ribosomal Subunit (2000) · cites this paper as De Stasio and Dahlberg 1990
In the citing paper
“It has been shown that loss of the ability of HygB to bind the 30S is also caused by mutation of either G1491 or C1409, which form a Watson-Crick base pair (De Stasio and Dahlberg 1990”
From The Structural Basis for the Action of the Antibiotics Tetracycline, Pactamycin, and Hygromycin B on the 30S Ribosomal Subunit (2000) · cites this paper as De Stasio and Dahlberg 1990
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Cited by
Brodersen, D. E., Clemons, W. M., Carter, A. P., Morgan-Warren, R. J., Wimberly, B. T., & Ramakrishnan, V. (2000) · Cell · doi:10.1016/s0092-8674(00)00216-6 · Cited by 950
“…has been shown that loss of the ability of HygB to bind the 30S is also caused by mutation of either G1491 with kinetic experiments that suggest that tetracycline only affects the rate of binding of EF-Tu ternary complex or C1409, which form a Watson-Crick base pair (De Stasio and Dahlberg, 1990).”
Background
Metadata
- Authors
- De Stasio, E. A., & Dahlberg, A. E.
- Venue
- Journal of Molecular Biology
- Publisher
- Elsevier BV
- Published
- 1990-03
- Volume
- 212
- Issue
- 1
- Pages
- 127-133
- ISSN
- 0022-2836 (ISSN-L)
- OpenAlex
- W1989241866
- Cited by
- 90
- References
- 0
- Citations by year
- 2021: 1 · 2019: 1 · 2015: 1 · 2014: 3 · 2013: 1 · 2012: 3
- FWCI
- 1.28
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of General Medical Sciences (GM19756, T32GM007601)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
- Keywords
- 16S rRNA, Escherichia coli, rRNA mutations, ribosome structure, 70S ribosome, aminoglycoside resistance, neomycin, kanamycin, gentamicin, apramycin, hygromycin, reversion, growth rate, rRNA secondary structure
- MeSH terms
- Anti-Bacterial Agents, Base Composition, Base Sequence, DNA, Bacterial, DNA, Ribosomal, Drug Resistance, Microbial, Escherichia coli, Molecular Sequence Data, Mutation, Nucleic Acid Conformation, Paromomycin, Phenotype, RNA, Bacterial, RNA, Ribosomal, RNA, Ribosomal, 16S
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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