Summary
The breakage of closed covalent λ DNA molecules in lysogenic host cells after ultravilet irradiation was investigated. In repair proficient host cells incisions are introduced immediately following irradiation. A steady-state of strand interruptions is observed within 20–50 seconds after irradiation, where the number of broken molecules is dose dependent for doses up to 600 ergs/mm2. No ultraviolet promoted strand breaks were observed in uvrA or uvrB lysogens, in accordance with previous results obtained by Shimada, Ogawa & Tomizawa [Molec. gen. Genet. 101, 245 (1968)]. In contrast, uvrC mutants have the ability to form breaks in superinfecting λ DNA molecules after ultraviolet irradiation. The ultraviolet specific endonucleolytic activity observed in uvrC host cells differs from that observed in uvr + host cells in that, (i) the first break is introduced at least 15 times slower, (ii) for doses below 300 ergs/mm2 the number of strand breaks is higher, (iii) the dose dependence terminates at a lower dose. The possible function of the uvrC gene product in the repair is discussed.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beukers, R., Ijlstra, J., Berends, W.: The effect of ultraviolet light on some components of the nucleic acids. IV. The origin of the U. V. sensitivity of deoxyribonucleic acid. Rec. Trav. chim. Pays-Bas 79, 101–104 (1960).
Boyce, R. P., Howard-Flanders, P.: Release of ultraviolet light-induced thymine dimers from DNA in E. coli K-12. Proc. nat. Acad. Sci. (Wash.) 51, 293–300 (1964).
Boyce, R. P., Kraiselburd, E., Ryan, S., Chessin, H.: Ring opening of covalent λ phage DNA circles after thermal induction of superinfected lysogens: Control by gene N. Virology 37, 679–680 (1969).
Boyle, J. R., Setlow, R. B.: Correlations between host-cell reactivation, ultraviolet reactivation and pyrimidine dimer excision in the DNA of bacteriophage λ. J. molec. Biol. 51, 131–144 (1970).
Ganesan, A. K., Smith, K. C.: Recovery of recombination deficient mutants of Escherichia coli K-12 from ultraviolet irradiation. Cold. Spr. Harb. Symp. quant. Biol. 33, 235–242 (1968).
Harm, W.: Mutants of phage T4 with increased sensitivity to ultraviolet. Virology 19, 66–71 (1963).
Howard-Flanders, P.: DNA-repair. Ann. Rev. Biochem. 37, 175–200 (1968).
Howard-Flanders, P., Boyce, R. P., Theriot, L.: Three loci in Escherichia coli K-12 that control the excision of pyrimidine dimers and certain other mutagen products from DNA. Genetics 53, 1119–1136 (1966).
Howard-Flanders, P., Wilkins, B. M., Rupp, W. D.: Genetic recombination induced by ultraviolet light. In: Molecular genetics (eds. Wittmann, H. G., Schuster, H.), p. 161–173. Berlin-Heidelberg-New York: Springer 1968.
Johansen, I., Gurvin, I., Rupp, W. D.: The formation of single-strand breaks in intracellular DNA by X-rays. Radiat. Res. 48, 599–612 (1971).
Kaplan, J. C., Kushner, S. R., Grossman, L.: Enzymatic repair of DNA. III. Properties of the UV-endonuclease and UV-exonuclease. Biochemistry 10, 3315–3324 (1971).
Kato, T.: Excision repair characteristics of recB − res −and uvrC − strains of Escherichia coli. J. Bact. 112, 1237–1246 (1972).
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 (Lond.) 224, 495–501 (1969).
Kushner, S. R., Kaplan, J. C., Ono, H., Grossman, L.: Enzymatic repair of deoxyribonucleic acid. IV. Mechanism of photoproduct excision. Biochemistry 10, 3325–3334 (1971).
Lucia, P. de, Cairns, J.: Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature (Lond.) 224, 1164–1166 (1969).
Nakayama, H., Okubo, S., Takagi, Y.: Repair of ultraviolet-damaged DNA in Micrococcus lysodeikticus. I. An endonuclease specific for ultraviolet-irradiated DNA. Biochim. biophys. Acta (Amst.) 228, 67–82 (1971).
Putte, P. van de, Sluis, C. A. van, Dillewijn, J. van, Rørsch, A.: The location of genes controlling radiation sensitivity in Escherichia coli. Mutation Res. 2, 97–110 (1965).
Rupp, W. D., Howard-Flanders, P.: Discontinuities in the DNA synthesized in an excision-defective strain of Escherichia coli following ultraviolet irradiation. J. molec. Biol. 31, 291–304 (1968).
Setlow, R. B.: The photochemistry, photobiology, and repair of polynucleotides. Progr. Nucl. Acid. Res. 8, 257–295 (1968).
Setlow, R. B., Carrier, W. L.: The disappearance of thymine dimers from DNA: An error-correcting mechanism. Proc. nat. Acad. Sci. (Wash.) 51, 226–231 (1964).
Setlow, R. B., Carrier, W. L., Bollum, F. J.: Pyrimidine dimers in UV-irradiation poly dI:dC. Proc. nat. Acad. Sci. (Wash.) 53, 1111–1118 (1965).
Shimada, K., Ogawa, H., Tomizawa, J.: Studies on radiation-sensitive mutants of E. coli. II. Breakage and repair of ultraviolet irradiated intracellular DNA of phage lambda. Molec. gen. Genet. 101, 245–256 (1968).
Taketo, A., Yasuda, S., Sekiguchi, M.: Initial step of excision repair in Escherichia coli: Replacement of defective function of uvr mutants by T4 endonuclease V. J. molec. Biol. 70, 1–14 (1972).
Wehrli, W., Staehelin, M.: Actions of the rifamycins. Bact. Rev. 35, 290–309 (1971).
Wulff, D. L., Rupert, C. S.: Disappearance of thymine photodimer in ultraviolet-irradiated DNA upon treatment with a photoreactivating enzyme from Baker's Yeast. Biochem. biophys. Res. Commun. 7, 237–240 (1962).
Yasuda, S., Sekiguchi, M.: Mechanism of repair of DNA in bacteriophage. II. Inability of ultraviolet-sensitive strains of bacteriophage in inducing an enzyme activity to excise pyrimidine dimers. J. molec. Biol. 47, 243–255 (1970).
Author information
Authors and Affiliations
Additional information
Communicated by G. Bertani
Rights and permissions
About this article
Cite this article
Seeberg, E., Johansen, I. Incisions in ultraviolet irradiated circular bacteriophage λ DNA molecules in excision proficient and deficient lysogens of E. coli . Molec. Gen. Genet. 123, 173–184 (1973). https://doi.org/10.1007/BF00267333
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00267333