Summary
We have found that the cells possessing the polA6 mutation affecting DNA polymerase I are unable to accept another mutation (uvr502) leading to UV-sensitivity. The introduction of the polA12 mutation determining the synthesis of a temperature sensitive DNA polymerase I into the uvr502 mutant results in the temperature sensitivity of colony forming ability of the double mutant. These data show that the uvr502 derivatives lacking DNA polymerase I are inviable. Reversions to temperature resistance in the population of the double mutant uvr502 polA12 may occur because of reverse mutations at one of the mutated sites or because of mutations suppressing DNA polymerase I deficiency but not UV- or MMS-sensitivity of revertants. DNA and protein synthesis in uvr502 polA12 cells continues after a shift to 45°C with rates almost indistinguishable from those in single mutants or wild type cells. No differences in DNA degradation were observed during incubation of single and double mutants at 45°C. The single strand molecular weight distribution of parent DNA from the double mutant as well as that from wild type cells is not affected by the shift to 45°C and 3 hours incubation at this temperature. We suggest that DNA polymerase I and/or the product altered by the uvr502 mutation are required for some step(s) of discontinuous DNA replication nonessential for the formation of acid insoluble DNA. The DNA polymerase I and the uvr gene product seem to be able to substitute for each other in accomplishing this process.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Cooper, P. K., Hanawalt, P. C.: Role of DNA polymerase I and REC system in excision repair in Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 65, 1156–1160 (1972).
Davis, B. D., Mingioli, E. S.: Mutants of Escherichia coli requiring methionine or vitamin B12. J. Bact. 60, 17–28 (1950).
Ganesan, A. T.: Studies on in vivo replication of Bacillus subtilis DNA. Cold Spr. Harb. Symp. quant. Biol. 33, 45–57 (1968).
Gellert, M., Little, J. W., Oshinsky, C. K., Zimmerman, S. B.: Joining of DNA strands by DNA ligase of E. coli. Cold Spr. Harb. Symp. quant. Biol. 33, 21–26 (1968).
Gross, J. D., Grunstein, J., Witkin, E. M.: Inviability of a recA - derivatives mutant of de Lucia and Cairns. J. molec. Biol. 58, 631–634 (1971).
Hanawalt, P. C.: Repair replication in the bacterial genome. In: Genetical aspects of radiosensitivity: Mechanisms of repair, p. 97. Vienna: International Atomic Energy Agency 1966.
Hanawalt, P. C., Pettijohn, D. E., Pauling, E. C., Brunk, C. F., Smith, D. W., Kanner, L. C., Couch, J. L.: Repair replication of DNA in vivo. Cold Spr. Harb. Symp. quant. Biol. 33, 187–194 (1968).
Kanner, L., Hanawalt, P. C.: Repair deficiency in a bacterial mutant defective in DNA polymerase. Biochem. biophys. Res. Commun. 39, 149–156 (1970).
Kelley, W. S., Whitfield, H. J.: Purification of an altered DNA polymerase from an Escherichia coli strain with a pol mutation. Nature (Lond.) 230, 33–36 (1971).
Kelly, R. B., Atkinson, M. R., Huberman, J. A., Kornberg, A.: Excision of thymine dimers an other mismatched sequences by DNA polymerase of Escherichia coli. Nature (Lond.) 224, 495–501 (1969).
Kelly, R. B., Cozzarelli, N. R., Deutscher, M. P., Lehman, I. R., Kornberg, A.: Enzymatic synthesis of deoxyribonucleic acid. XXXII. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break. J. biol. Chem. 245, 39–45 (1970).
Kuempel, P. L., Veomett, G. E.: A possible function of DNA polymerase in chromosome replication. Biochem. biophys. Res. Commun. 41, 973–980 (1970).
Masamune, Y., Fleischman, R. A., Richardson, C. C.: Enzymatic removal and replacement of nucleotides at a single strand break in deoxyribonucleic acid. J. biol. Chem. 246, 2680–2691 (1971).
Masamune, Y., Richardson, C. C.: Strand displacement during deoxyribonucleic acid synthesis at a single strand breaks. J. biol. Chem. 246, 2692–2701 (1971).
Mattern, I. E.: A class of UV-sensitive mutants of E. coli K12 that show a high spontaneous mutation frequency. In: First Europ. Biophys. Congr., Baden, Austria, 1971, p. 237–240.
Monk, M., Kinross, J.: Conditional lethality of recA and recB derivatives of a strain of E. coli K-12 with a temperature sensitive deoxyribonucleic acid polymerase. J. Bact. 109, 971–978 (1972).
Monk, M., Peacey, M., Gross, J. D.: Repair of damage induced by ultraviolet light in DNA polymerase defective E. coli cells. J. molec. Biol. 58, 623–630 (1971).
Okazaki, R., Arisuwu, M., Sugino, A.: Slow joining of newly replicated DNA chains in DNA polymerase I deficient E. coli mutants. Proc. nat. Acad. Sci. (Wash.) 68, 2954–2957 (1971).
Okazaki, R., Okazaki, T., Sakebe, K., Sugimoto, K., Kainuma, R., Sugino, A., Iwatsuki, N.: In vivo mechanism of DNA chain growth. Cold Spr. Harb. Symp. quant. Biol. 33, 129–143 (1968).
Paterson, M. C., Boyle, J. M., Setlow, R. B.: Ultraviolet and X-ray-induced responses of a deoxyribonucleic acid polymerase-deficient mutant of Escherichia coli. J. Bact. 107, 61–67 (1971).
Putte, P. van de, Sluis, C. A. van, Dillewijn, Y. van, Rörsch, A.: The location of genes controlling radiation sensitivity in E. coli. Mutation Res. 2, 97–110 (1965).
Sinzinis, B. I., Smirnov, G. B., Saenko, A. S.: Effect of polA1 mutation on the ability of Escherichia coli K-12 to repair single strand breaks of DNA induced by methyl methanesulfonate. Biochim. biophys. Acta (Amst.) 247, 635–639 (1971).
Smirnov, G. B., Filkova, E. V., Skavronskaya, A. G.: The mutator propenty of uvr502 mutation affecting UV-sensitivity of Escherichia coli. Molec. gen. Genet. 118, 51–56 (1972).
Smirnov, G. B., Skavronskaya, A. G.: The lethal and mutagenic action of UV light and mitomycin C on the mutants of Escherichia coli with reduced dark repair ability. Genetika (USSR) 4, 105–110 (1968).
Smirnov, G. B., Skavronskaya, A. G.: Location of uvr502 mutation on the chromosome of Escherichia coli K-12. Molec. gen. Genet. 113, 217–221 (1971).
Town, C. D., Smith, K. C., Kaplan, H. S.: DNA polymerase required for rapid repair of X-ray-induced DNA-strand breaks in vivo. Science 172, 851–854 (1971).
Willetts, N. S., Clark, A. J.: Characteristics of some multiply recombination-deficient strains of Escherichia coli. J. Bact. 100, 231–239 (1969).
Author information
Authors and Affiliations
Additional information
Communicated by H. Böhme
Rights and permissions
About this article
Cite this article
Smirnov, G.B., Filkova, E.V., Skavronskaya, A.G. et al. Loss and restoration of viability of E. coli due to combinations of mutations affecting DNA polymerase I and repair activities. Molec. Gen. Genet. 121, 139–150 (1973). https://doi.org/10.1007/BF00277528
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00277528