Unusual growth properties of a bacterial strain lacking DNA polymerase

doi:10.1016/S0006-291X(71)80117-1

Biochemical and Biophysical Research Communications

Volume 44, Issue 3, 6 August 1971, Pages 546-549

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Summary

The DNA polymerase-deficient E. coli strain pol A₁⁻ exhibits a 5% plating efficiency when grown on synthetic liquid medium and plated on a nutritionally rich solid medium. This phenomenon is not seen when the composition of the liquid and solid media is identical. Accordingly in order to obtain meaningful results with this strain, the growth conditions must be controlled carefully.

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Cited by (10)

Evaluation of a DNA polymerase-deficient mutant of E. coli for the rapid detection of carcinogens
1976, Chemico-Biological Interactions
Differential growth inhibition of two E. coli cultures was evaluated as a rapid screening technique for chemical carcinogens. Of the carcinogens tested, only “direct acting” carcinogens produced positive results. Furthermore, this test is not a quantitative assay in that neither was a dose—response relationship seen nor did potent carcinogens necessarily show a greater response than weaker carcinogens.
Most of the carcinogens tested are considered to require metabolic activation in order to exert their carcinogenic action. Despite many attempts, including several variations of reaction conditions, metabolic activation by rat liver fractions was not apparent. Many of these carcinogens are insoluble in water and may not diffuse through the agar and therefore not reach the indicator organism.
A number of chemicals that are not carcinogenic produced positive results with this assay. Many of these substances are oxidants or oxidation products which are highly reactive with DNA as well as with other cellular constituents. Therefore, it is possible that the toxicity exhibited by these chemicals was caused by a reaction with some essential cellular constituent other than DNA and such damage would not be repairable by DNA polymerase. These observations limit the usefulness of the P3478 E. coli technique in its present form as a prescreen for chemical carcinogens.
Hydroxylamine-O-sulfonic acid: In vitro and possible in vivo reaction with DNA
1973, Chemico-Biological Interactions
Exposure of DNA solutions to low levels (2 · 10⁻³M) of hydroxylamine-O-sulfonic acid (HOS) resulted in limited degradation accompanied by increases in the buoyant density of the DNA. The thermal helix-coil profile of the DNA was not changed significantly following exposure to HOS. Upon thermal denaturation, the treated DNA specimens exhibited buoyant density values similar to those of heated control DNA. This is taken to mean that an added function or a modified DNA base is removed upon heating. Exposure of DNA to elevated levels of HOS (>0.2 M) resulted in extensive degradation which was accompanied by spectral changes: a hyperchromic shift and an increase in the wavelength of maximum absorbance.
Exposure of individual deoxynucleosides to HOS also resulted in spectral changes and in the detection of new reaction products by paper Chromatographic means.
HOS preferentially inhibited the growth of a bacterial strain deficient in DNA polymerase. This is a property also exhibited by known mutagens and carcinogens. This is taken to mean that HOS is capable of reacting with the DNA of living cells as well.
Sodium hypochlorite and sodium perborate: preferential inhibitors of DNA polymerase-deficient bacteria
1973, Mutation Research/Environmental Mutagenesis and Related Subjects
Effect of the recC gene in Escherichia coli on frequencies of ultraviolet-induced mutants
1973, Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
UV induction of Lac⁻ mutations was compared with UV induction of Mal⁺ mutations in E. coli B/r strains differing in the recC gene. The frequency of Lac⁻ mutants per survivor induced by the same dose was not significantly affected by the recC gene but the percentage of pure rather than sectored Lac⁻ colonies was greater when the recC gene was present. On the other hand, as reported previously, frequencies of Mal⁺ mutants induced by the same UV dose were lower when the strain was recC. The reduction factor was the same as for spontaneous Mal⁺ mutants. The difference in the effect of the recC gene on the yields of Lac⁻ and Mal⁺ mutants can be explained by taking into account the influence of lethal sectoring, which introduces an artifact when mutants arising in the recC strain are scored selectively as in the case for Mal⁺ mutants, but not when the scoring is non-selective as for Lac⁻ mutants. Lethal sectoring as indicated by a discrepancy between total cell counts and numbers of colony-formers, was observed for the recC strain growing in liquid minimal medium corresponding to the agar medium used to score Mal⁺ mutants but was not observed for the rec⁺ strain. Both strains showed lethal sectoring in the liquid medium corresponding to the agar medium to score Lac⁻ mutants. The hypothesis concerning the role of lethal sectoring in the selective scoring of mutants arising in a recC background is supported by evidence concerning the UV induction of mutants in a polA1 background. Like the recC gene, the polA1 gene did not affect yields of Lac⁻ mutants. However, unlike the recC gene, the polA1 gene has previously been shown not to influence UV yields of prototrophic mutations (scored selectively) and not to cause lethal sectoring except under irrelevant conditions.
DNA synthesis in prokaryotes: Replication
1973, Progress in Biophysics and Molecular Biology
Peroxide sensitivity of cold‐shocked Salmonella typhimurium and Escherichia coli and its relationship to minimal medium recovery
1986, Journal of Applied Bacteriology

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Unusual growth properties of a bacterial strain lacking DNA polymerase

Summary

Biochem. Biophys. Res. Comm.

Biochem. Biophys. Res. Comm.

Nature

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Nature

Nature

Nature

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