Summary
The influence of UV-specific endonuclease and medium composition on the frequency and spectrum of genic mutations in Escherichia coli KI2 uvr + (with normal repair enzymes) and urv A6 (defective in UV-specific endonuclease) was studied. Mutations at the locus glu (gene controlling assimilation of glucose) were induced by ultra-violet irradiation and hydroxylamine treatment. To identify mutant colonies, triphenyl tetrazolium chloride (TTC) was added to the medium since it coloured the mutant colonies bright crimson and readily permitted distinction between pure mutant clones (complete mutations) and mixed clones (mosaic or sector mutations).
A maximum mutation frequency after UV-irradiation was observed in E. coli uvr + cells but not in the E. coli uvr A6 strain. The curve of mutagenesis with a maximum was found in both studied strains after treatment by hydroxylamine which did not cause DNA damage recognized by UV-specific endonuclease.
The highest frequency of mutations (at the point of maximum) in the series of experiments with enriched growth medium was almost 10 times higher than in the series of the experiments with poor medium.
It was established that in bacteria with normal repair enzymes the frequency of complete mutations was higher than the frequency of mosaic mutations. It was also observed that the rate of UV-mutagenesis was higher in the case of E. coli uvr +.
The study of the distribution of mosaic mutant sectors in experiments with bacteria suspended in either a nutrient broth or a buffer during UV-irradiation revealed that the size of mutant sectors was rather variable and that the differences in the number of nucleoids per cell did not always determine the distribution of mutant sector sizes.
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Abbreviations
- HA:
-
Hydroxylamine hydrochloride
- TTC:
-
Triphenyl tetrazolium chloride
- TCA:
-
Trichloroacetic acid
Literature
Carrier, W.L.; Setlow, R.B.: Endonucleases from Micrococcus luteus which has activity toward ultraviolet-irradiated deoxyribonucleic acid: purification and properties. J. Bacteriol. 102, 178–186 (1970)
Cole, R.S.; Levitan, D.; Sinden, R.R.: Removal of psoralen interstrand crosslinks from DNA of Escherichia coli: mechanism and genetic control. J. Mol. Biol. 103, 39–59 (1976)
Dubinin, N.P.; Soyfer, V.N.: Chromosome breakage and complete genic mutation production in molecular terms. Mutation Res. 8, 353–365 (1969)
Helmstetter, C.E.; Cummings, D.J.: Bacterial synchronization by selection of cells at division. Proc. Natl. Acad. Sci. (USA) 50 b, 767–773 (1963)
Kelly, R.B.; Atkinson, M.R.; Huberman, J.A.; Kornberg, A.: Excision of thymine dimers and other mismatched sequences by DNA polymerase of Escherichia coli. Nature 244, 495–501 (1969)
Kushner, S.R.; Kaplan, I.C.; Ono, H.; Grossman, L.: Enzymic repair of deoxyribonucleic acid. IV. Mechanism of photoproduct excision. Biochemistry 10, 3325–3334 (1971)
Lieberman, M.W.; Poirier, M.C.: Base pairing and template specificity during deoxyribonucleic acid repair synthesis in human and mouse cells. Biochemistry 13, 5384–5388 (1974)
Makino, R.; Okada, S.: Comparative studies of the effects of carcino-genic and antitumor agents on the DNA replication of cultured mammalian cells. Mutation Res. 23, 387–394 (1974)
Nischioka, H.; Doudney, C.O.: Different modes of loss of photoreversibility of mutation and lethal damage in ultraviolet-light resistant and sensitive bacteria. Mutation Res. 8, 215–228 (1969)
Nischioka, H.; Doudney, C.O.: Different modes of loss of photoreversibility of ultraviolet-light induced true and supressor mutations to tryptophan independence in an auxotrophic strain of E. coli. Mutation Res. 9, 349–358 (1970)
Ogur, M.; John, R.; Nagai, S.: Tetrazolium overlay technique for population studies of respiration deficiency in yeast. Science 125, 928–929 (1957)
Paribok, V.P.; Tomilin, N.V.: Recognition of pyrimidine dimers in DNA by the incision enzyme from Micrococcus lysodeikticus. Nature 230, 210–211 (1971)
Paul, C.; Taylor, R.; Friedberg, E.C.: Kinetics of thymine dimer excision from phage T4 DNA in vivo. Biophys. J. 16 (2) part 2, pp. 58 (1976)
Roberts, J.J.; Sturrock, J.F.; Ward, K.N.: Enchancement by caffeine of alkylation induced cell death, mutations and chromosome aberrations in Chinese hamster cells as result of inhibition of postreplicative repair. Mutation Res. 26, 129–143 (1974)
Sedgwick, S.G.: Misrepair of overlapping daughter strand gaps as a possible mechanism for UV-induced mutagenesis in uvr strains of Escherichia coli: a general model for induced mutagenesis by misrepair (SOS repair) of closely spaced DNA lesions. Mutation Res. 41, 185–200 (1976)
Sedgwick, S.G.; Bridges, B.A.: Survival, mutation and capacity to repair single-strand DNA breaks after gamma irradiation in different exr − strains of E. coli. Molec. Gen. Genet. 119, 93–102 (1972)
Simon, T.J.; Smith, C.A.; Friedberg, F.C.: Action of bacteriophage T4 ultraviolet endonuclease on duplex DNA containing one ultra-violet-irradiated strand. J. Biol. Chem. 250, 8748–8752 (1975)
Smith, K.C.: The inhibition of DNA repair processes. Proc. XI. Intern. Cancer Congr. 5, 105–110 (1975)
Soyfer, V.N.: The Molecular Mechanisms of Mutagenesis. (in Russian) Moscow: Publ. House ‘Nauka’ 1969
Soyfer, V.N.: Neueste Forschungsergebnisse und Perspektiven der Mutationstheorie. Moderne Medizin 1, 134–141 (1971)
Soyfer, V.N.: Participation of the intracellular enzymes in the control of mutation process. In: Molecular mechanisms of genetic processes. (in Russian) pp 364–377. Moscow: 1972a
Soyfer, V.N.: The role of the DNA molecules conformation in the initiation of dark repair and possible consequences of ‘conformational mistakes’ of the repair systems. In: Abstr. IV. intern. biophys. congr., Sections V-VII. pp. 320–321 (1972b)
Soyfer, V.N.; Kartel, N.A.: Participation of the intracellular enzymes in the control of mutation process. Part 2: Influence of inhibition of repair and replication on X-ray induced chromosomal aberrations in barley. Theor. Appl. Genet. 53, 9–15 (1978)
Soyfer, V.N.; Titov, Yu.B.; Kartel, N.A.; Sen', L.A.: The role of the mistakes of repair enzymes in the induction of genic and chromosome mutations. Bull. Acad. Sci. USSR Biol. 3, 363–373 (1977)
Titov, Yu.B.; Soyfer, V.N.: Differences in the inhibition of cell division and DNA synthesis by ultraviolet irradiation of E. coli uvr + and uvr A6. Effects of hydroxyurea on DNA synthesis in irradiated cells. Mol. Biol. 11, 84–91 (1977) (transl. by Plenum Publ. Co.)
Town, C.D.; Smith, K.C.; Kaplan, H.S.: Repair of X-ray damage to bacterial DNA. Current topics Radiat. Res. 8, 351–399 (1973)
Verly, W.G.: Monofunctional alkylating agents and apurinic sites in DNA. Biochem. Pharm. 23, 3–8 (1974)
Witkin, E.M.: Mutation-proof and mutation-prone modes of survival in derivatives of Escherichia coli B differing in sensitivity to UV-light. Brookhaven Symp. Biol. 20, 17–55 (1967)
Witkin, E.M.: Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli Bacter. Rev. 40, 869–907 (1976)
Youngs, D.A.; Smith, K.C.: X-ray sensitivity and repair capacity of a pol AI exr A strain of E. coli KI2. J. Bacter. 114, 121–127 (1973)
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Communicated by D.K. Belyaev
Other papers of this series are: Soyfer 1972; Soyfer et al. 1977; Soyfer and Kartel 1978
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Soyfer, V.N., Titov, Y.B. Participation of the intracellular enzymes in the control of mutational processes. Theoret. Appl. Genetics 55, 119–128 (1979). https://doi.org/10.1007/BF00295437
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DOI: https://doi.org/10.1007/BF00295437