Massive overproduction of dihydrofolate reductase in bacteria as a response to the use of trimethoprim
Open accessAbstract
Among several observations of greatly increased levels of chromosomal dihydrofolate reductase as a cause of resistance to high concentrations of the antifolate drug trimethoprim, in clinically isolated bacteria, one is described here of a strain of Escherichia coli overproducing dihydrofolate reductase several hundredfold. The chromosomally located resistance gene of this strain was isolated, inserted into a plasmid vector, and analyzed for its nucleotide sequence. The structural gene for the overproduced dihydrofolate reductase was found to be identical to that of E. coli K12, with nine exceptions, of which seven resulted in synonymous codon usage. Two transversions resulted in a substitution of Gly for Trp at amino acid position 30, and of Gln for Glu at position 154. Six of the nine base changes resulted in codons more frequently used. The Gly substitution which leads to a less commonly used codon, was thought to relate to the observed threefold increase in Ki for trimethoprim. Furthermore, a C→T transition was found in the –35 region of the promoter, increasing its homology with the E. coli consensus promoter sequence. In the ribosome‐binding area of the resistant strain, finally, seven base changes were observed, two of which resulted in a five‐base sequence of complementarity with the 3′‐end of ribosomal 16S RNA. The distance between the —10 site of the promoter and the start codon for translation was finally increased one base pair by the insertion of an A at position + 9 in the resistant strain. These genetic changes towards more efficient transcriptional and translational start sequences and towards increased mRNA expressivity are interpreted to reflect an evolutionary adaptation to the presence of antifolates.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Promoter mutations that increase production of DHFR or DHPS overwhelm the ability of TMP-SMX to inhibit folate production and permit bacterial survival.
“These mutations result in the production of increased quantities of the above enzymes, “overwhelming” the ability of TMP-SMX to inhibit folate production and permitting bacterial survival (82, 83).”
From Mechanisms of Antibiotic Resistance (2016) · cites this paper as [82]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
[57] Sequencing end-labeled DNA with base-specific chemical cleavages
Maxam, A. M., & Gilbert, W. (1980) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(80)65059-9 · Cited by 14,678 · see citing works
A complementation analysis of the restriction and modification of DNA in Escherichia coli
Boyer, H. W., & Roulland-Dussoix, D. (1969) · Journal of Molecular Biology · doi:10.1016/0022-2836(69)90288-5 · Cited by 3,847 · see citing works
REGULATORY SEQUENCES INVOLVED IN THE PROMOTION AND TERMINATION OF RNA TRANSCRIPTION
Rosenberg, M. J., & Court, D. (1979) · Annual Review of Genetics · doi:10.1146/annurev.ge.13.120179.001535 · Cited by 2,726 · see citing works
A graphics model building and refinement system for macromolecules
Jones, T. A. (1978) · Journal of Applied Crystallography · doi:10.1107/s0021889878013308 · Cited by 1,848 · see citing works
Determinant of cistron specificity in bacterial ribosomes
Shine, J., & Dalgarno, L. (1975) · Nature · doi:10.1038/254034a0 · Cited by 1,365 · see citing works
Codon usage in bacteria: correlation with gene expressivity.
Gouy, M., & Gautier, C. (1982) · Nucleic Acids Research · doi:10.1093/nar/10.22.7055 · Cited by 1,182 · see citing works
Alt, F. W., Kellems, R. E., Bertino, J. R., & SCHIMKE, R. T. (1978) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)34875-5 · Cited by 934 · see citing works
Ikemura, T. (1981) · Journal of Molecular Biology · doi:10.1016/0022-2836(81)90363-6 · Cited by 875 · see citing works
Sequences at the somatic recombination sites of immunoglobulin light-chain genes
Sakano, H., Huppi, K. E., Heinrich, G., & Tonegawa, S. (1979) · Nature · doi:10.1038/280288a0 · Cited by 857 · see citing works
Advanced Bacterial Genetics: A Manual for Genetic Engineering
Davis, R. W., Botstein, D., & Roth, J. R. (1980) · Medical Entomology and Zoology · Cited by 823 · see citing works
Plasmid ColE1 as a Molecular Vehicle for Cloning and Amplification of DNA
Hershfield, V., Boyer, H. W., Yanofsky, C., et al. (1974) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.71.9.3455 · Cited by 542 · see citing works
Malke, H. (1981) · Zeitschrift für allgemeine Mikrobiologie · doi:10.1002/jobm.19810211013 · Cited by 413 · see citing works
Nucleotide sequence of the E. coli gene coding for dihydrofolate reductase
Smith, D. R., & Calvo, J. M. (1980) · Nucleic Acids Research · doi:10.1093/nar/8.10.2255 · Cited by 375 · see citing works
Dihydrofolate Reductase: X-ray Structure of the Binary Complex with Methotrexate
Matthews, D. A., Alden, R. A., Bolin, J. T., et al. (1977) · Science · doi:10.1126/science.17920 · Cited by 324 · see citing works
Nucleotide sequence of cro, cII and part of the O gene in phage λ DNA
Schwarz, E., Scherer, G., Hobom, G., & Kössel, H. (1978) · Nature · doi:10.1038/272410a0 · Cited by 222 · see citing works
Directed Mutagenesis of Dihydrofolate Reductase
Villafranca, J. E., Howell, E. E., Voet, D. H., et al. (1983) · Science · doi:10.1126/science.6356360 · Cited by 209 · see citing works
Spacer mutations in the lac ps promoter.
Stefano, J. E., & Gralla, J. D. (1982) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.79.4.1069 · Cited by 157 · see citing works
Evolution of antibiotic resistance gene function
Koch, A. L. (1981) · Microbiological Reviews · doi:10.1128/mr.45.2.355-378.1981 · Cited by 66 · see citing works
Smith, D. R., & Calvo, J. M. (1982) · Molecular and General Genetics MGG · doi:10.1007/bf00384386 · Cited by 44 · see citing works
Tennhammar-Ekman, B., & Sköld, O. (1979) · Plasmid · doi:10.1016/0147-619x(79)90017-9 · Cited by 39 · see citing works
Self-citationChang, A. C. Y., Erlich, H. A., Gunsalus, R. P., et al. (1980) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.77.3.1442 · Cited by 30 · see citing works
Flensburg, J., & Sköld, O. (1984) · Journal of Bacteriology · doi:10.1128/jb.159.1.184-190.1984 · Cited by 13 · see citing works
Self-citation
Flagged first, then by citations
Research works printed but not linked: 3
- Hershfield, V., Boyer, H. W., Yanofsky, C., Lovett, M. & Helinski, D. R. (1974) Proc. Natl Acad. Sci. USA 71, 3455- 3459.
- Stefano, J. E. & Gralla, J. D. (1982) Proc. Natl Acad. Sci. USA
- Chang, A. C. Y., Erlich, H. A,, Gunsalus, R. P., Nunberg, J. H., Kaufman, R. J., Schimke, R. T. & Cohen, S. (1980) Proc. Natl Acad. Sci. USA 77,1442 -1446.
Cited by
Sorted by citations
Mechanisms of Antibiotic Resistance
Munita, J. M., & Arias, C. A. (2016) · Microbiology Spectrum · doi:10.1128/microbiolspec.vmbf-0016-2015 · Cited by 2,782 · see citing works
Evolutionary paths to antibiotic resistance under dynamically sustained drug selection
Toprak, E., Veres, A., Michel, J.-B., Chait, R., Hartl, D. L., & Kishony, R. (2012) · Nature Genetics · doi:10.1038/ng.1034 · Cited by 764 · see citing works
Mechanisms of antibiotic resistance
Munita, J. M., & Arias, C. A. (2016) · Virulence Mechanisms of Bacterial Pathogens · doi:10.1128/9781555819286.ch17 · Cited by 421 · see citing works
- Authors
- FLENSBURG, J., & SKÖLD, O.
- Affiliations
- Uppsala University (SE)
- Corresponding author
- SKÖLD, Ola
- Venue
- European Journal of Biochemistry
- Publisher
- Wiley
- Published
- 1987-02
- Volume
- 162
- Issue
- 3
- Pages
- 473-476
- ISSN
- 0014-2956 (ISSN-L)
- 1432-1033
- OpenAlex
- W2033843473
- Cited by
- 103
- References
- 22
- Citations by year
- 2026: 5 · 2025: 2 · 2024: 10 · 2023: 1 · 2022: 2 · 2021: 6 · 2020: 7 · 2019: 6 · 2018: 7 · 2017: 5 · 2016: 4 · 2015: 2 · 2014: 4 · 2013: 1 · 2012: 3
- FWCI
- 2.07
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- European Journal of Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Academy of Pharmaceutical Sciences
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Antibiotic Resistance in Bacteria
- Health Sciences › Medicine › Infectious Diseases › Antimicrobial Resistance in Staphylococcus
- Keywords
- dihydrofolate reductase, trimethoprim, Escherichia coli, antibiotic resistance, chromosomal resistance genes, synonymous codon usage, promoter mutations, ribosome binding site, translation initiation sites, antifolate, drug resistance evolution
- MeSH terms
- Base Sequence, Chromosomes, Bacterial, Drug Resistance, Microbial, Escherichia coli, Genes, Promoter Regions, Genetic, Tetrahydrofolate Dehydrogenase, Trimethoprim
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
