PMID:7032590

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Citation

Muir, ME, Hanwell, DR and Wallace, BJ (1981) Characterization of a respiratory mutant of Escherichia coli with reduced uptake of aminoglycoside antibiotics.Biochim. Biophys. Acta 638:234-41

Abstract

A strain of Escherichia coli (NSW77) which is partially resistant to streptomycin was isolated by selecting for growth on plates supplemented with 12.5 micrograms/ml streptomycin, a concentration which completely inhibits growth of wild-type strains. The low-level resistance of the mutant appears to result from a reduced ability to accumulate streptomycin intracellularly. In addition, the mutant strain is unable to use succinate for growth because of a defective respiratory chain. Thus, membranes of the mutant strain were found to have approximately half the NADH and D-lactate oxidase activity of the parent strain. Moreover, membranes of the mutant were found to contain demethyl-menaquinone and, in place of ubiquinone, a structural analogue, 2-octaprenyl-3-methyl-6-methoxy-1,4 benzoquinone. The mutation responsible for both the Suc-phenotype and partial resistance to streptomycin was found to be located near minute 15 on the bacterial chromosome. Both the biochemical and genetic evidence suggests the the mutation in strain NSW77 resides in the ubi F gene. Another previously characterized ubi F strain was also found to have a reduced capacity to take up an aminoglycoside antibiotic (gentamicin). These results suggest that the respiratory defects in ubi F strains are responsible for the reduced capacity of such strains to accumulate aminoglycosides.

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Keywords

Biological Transport; Drug Resistance, Microbial; Escherichia coli; Kinetics; Mixed Function Oxygenases; Multienzyme Complexes; Mutation; NADH, NADPH Oxidoreductases; Oxidoreductases; Oxygen Consumption; Proline; Species Specificity; Streptomycin; Succinates

Main Points of the Paper

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Materials and Methods Used

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Phenotype Annotations

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Phenotype of Taxon Information Genotype Information (if known) Condition Information OMP ID OMP Term Name ECO ID ECO Term Name Notes Status

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JP2140
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: ubiF+
  • Genotype of Experimental Strain : ubiF mutation in NSW77
  • Reference Condition:

0000181

in vitro culture assay data

Partial streptomycin resistance is conferred due to a mutation in the ubiF gene, resulting in reduced uptake of streptomycin. See Figure 1 for full results.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JP2140
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: ubiF+
  • Genotype of Experimental Strain : ubiF mutation in NSW77
  • Reference Condition:

0000252

metabolite accumulation

0000181

in vitro culture assay data

ubiF strain NSW77 had a higher rate of accumlation of the amino acid Proline than the parent strain or wild type. See figure 3 for full proline experimental results.

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: JP2140
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: ubiF+
  • Genotype of Experimental Strain : ubiF mutation in NSW77
  • Reference Condition:

0000220

oxidase phenotype

0000181

in vitro culture assay data

The ubiF strain has reduced oxidase activity in regards to the Parent strain. NADH and D-Lactose oxidase was reduced to 45 percent and Succinic oxidase 7 percent of the wild type strain. See table 1 of the experiment for the full results.


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