Summary
We constructed a strain of E. coli K12 carrying polA1 (an amber mutation of the DNA polymerase I gene; De Lucia and Cairns, 1969), and sup-126 (a temperature-sensitive amber suppressor; Nagata and Horiuchi, 1973). DNA polymerizing activity of the enzyme in this strain is virtually undetectable if the cells are grown at 42°C, but if grown at 30°C it is sufficiently present. By mutagenizing this strain, and after appropriate screening, we obtained mutants no longer able to grow at 42°, but able to do so when the normally functioning polA gene is present. One of them, called TS41, was most extensively studied. It acquired a mutation named pdeB41 which was found to be located between ilv and metE on the E. coli linkage map. Its phenotype is pleiotropic. The mutation by itself, i.e., if present in a polA + cell, does not kill the cell at 42°, but does so as in TS41 when it is reconstructed into a pdeB41 polA1 sup-126 triple mutant by P1 transduction. The mutation by itself renders the cell sensitive to UV, and tolerant to phage λ deficient in recombination. It is also a mutator.
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References
Baas, P. D., Jansz, H. S.: Asymmetric information transfer during Φ X 174 DNA replication. J. molec. Biol. 63, 557–568 (1972).
Brutlag, D., Kornberg, A.: Enzymatic synthesis of deoxyribonucleic acid. XXXVI. A proofreading function for the 3′→5′ exonuclease activity in deoxyribonucleic acid polymerases. J. biol. Chem. 247, 241–248 (1972).
Calendar, R., Lindqvist, B., Sironi, G., Clark, A. J.: Characterization of rep − mutants and their interaction with P2 phage. Virology 40, 72–83 (1970).
Clark, A. J.: Toward a metabolic interpretation of genetic recombination of E. coli and its phages. Ann. Rev. Microbiol. 25, 437–464 (1971).
De Lucia, P., Cairns, J.: Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature (Lond.) 224, 1164–1166 (1969).
Doerfler, W., Hogness, D. S.: Gene orientation in bacteriophage lambda as determined from the genetic activities of heteroduplex DNA formed in vitro. J. molec. Biol. 33, 661–678 (1968).
Eggertsson, G., Adelberg, E. A.: Map positions and specificities of suppressor mutation in Escherichia coli K-12. Genetics 52, 319–340 (1965).
Gefter, M. L., Hirota, Y., Kornberg, T., Wechsler, J. A., Barnoux, C.: Analysis of DNA polymerase II and III in mutants of Escherichia coli thermosensitive for DNA synthesis. Proc. nat. Acad. Sci. (Wash.) 68, 3150–3153 (1971).
Goebel, W.: Replication of the DNA of the colicinogenic factor E1 (col E1) at the restrictive temperature in a DNA replication mutant thermosensitive for DNA polymerase III. Nature (Lond.) New Biol. 237, 67–70 (1972).
Gross, J., Gross, M.: Genetic analysis of an E. coli strain with a mutation affecting DNA polymerase. Nature (Lond.) 224, 1166–1168 (1969).
Gross, J. D.: DNA replication in bacteria. In: Current topics in microbiology and immunology, vol. 57, p. 39–74. Berlin-Heidelberg-New York: Springer 1972.
Gross, J. D., Grunstein, J., Witkin, E. M.: Inviability of recA - derivatives of the DNA polymerase mutant of De Lucia and Cairns. J. molec. Biol. 58, 631–634 (1971).
Ikeda, H., Tomizawa, J.: Transducing fragments in generalized transduction by phage P1. I. Molecular origin of the fragments. J. molec. Biol. 14, 85–109 (1965).
Kingsbury, D. T., Helinski, D. R.: DNA polymerase as a requirement for the maintenance of the bacterial plasmid colicinogenic factor E1. Biochem. biophys. Res. Commun. 41, 1538–1544 (1971).
Kuempel, P. L., Veomett, G. E.: A possible function of DNA polymerase in chromosome replication. Biochem. biophys. Res. Commun. 41, 973–980 (1970).
Low, B.: Formation of merodiploids in matings with a class of rec - recipient strains of Escherichia coli K12. Proc. nat. Acad. Sci. (Wash.) 60, 160–167 (1968).
Manly, K. F., Signer, E. R., Radding, C. M.: Nonessential functions of bacteriophage λ. Virology 37, 177–188 (1969).
Monk, M., Kinross, J.: Conditional lethality of recA and recB derivatives of a strain of Escherichia coli K-12 with a temperature-sensitive deoxyribonucleic acid polymerase I. J. Bact. 109, 971–978 (1972).
Morse, D. E., Yanofsky, C.: Amber mutants of the rpR regulatory gene. J. molec. Biol. 44, 185–193 (1969).
Nagata, T., Horiuchi, T.: Isolation and characterization of a temperature-sensitive amber suppressor mutant of Escherichia coli K12. Molec. gen. Genet. 123, 77–88 (1973).
Ogawa, H., Shimada, K., Tomizawa, J.: Studies on radiation-sensitive mutants of E. coli. Molec. gen. Genet. 101, 227–244 (1968).
Okazaki, R., Arisawa, M., Sugino, A.: Slow joining of newly replicated DNA chains in DNA polymerase I-deficient Escherichia coli mutants. Proc. nat. Acad. Sci. (Wash.) 68, 2954–2957 (1971).
Okazaki, R., Sugino, A., Hirose, S., Okazaki, T., Imae, Y., Kainuma-Kuroda, R., Ogawa, T., Arisawa, M., Kurosawa, Y.: The discontinuous replication of DNA. In: R. D. Wells, R. B. Inman (eds.), in press. DNA synthesis in vitro. 2nd Steenbock Symp. Baltimore: University Park Press 1972.
Pittard, J., Loutit, J. S., Adelberg, E. A.: Gene transfer by F′ strains of Escherichia coli K-12. J. Bact. 85, 1394–1401 (1963).
Putte, P. van de, Sluis, C. A. van, Dillewijn, J. van, Rörsch, A.: The location of genes controlling radiation sensitivity in Escherichia coli. Mutation Res. 2, 97–110 (1965).
Setlow, P., Brutlag, D., Kornberg, A.: Deoxyribonucleic acid polymerase: Two distinct enzymes in one polypeptide. I. A proteolytic fragment containing the polymerase and 3′→5′ exonuclease functions. J. biol. Chem. 247, 224–231 (1972).
Setlow, P., Kornberg, A.: Deoxyribonucleic acid polymerase: Two distinct enzymes in one polypeptide. II. A proteolytic fragment containing the 5′→3′ exonuclease function. Restoration of intact enzyme functions from the two proteolytic fragments. J. biol. Chem. 247, 232–240 (1972).
Signer, E.: General recombination. In: A. D. Hershey (ed.), p. 139–174. The bacteriophage lambda. New York: Cold Spring Harbor Laboratory 1971.
Smirnov, G. B., Filkova, E. V., Skavronskaya, A. G.: The mutator property of uvr502 mutation affecting UV sensitivity of Escherichia coli. Molec. gen. Genet. 118, 51–56 (1972).
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).
Spatz, H. Ch., Trautner, T. A.: One way to do experiments on gene conversion? Transfection with heteroduplex SPP1 DNA. Molec. gen. Genet. 109, 84–106 (1970).
Taylor, A. L.: Current linkage map of Escherichia coli. Bact. Rev. 34, 155–175 (1970).
Taylor, A. L., Adelberg, E. A.: Linkage analysis with very high frequency males of Escherichia coli. Genetics 45, 1233–1243 (1960).
Zissler, J., Signer, E., Schaefer, F.: The role of recombination in growth of bacteriophage lambda. I. The gamma gene. In: A. D. Hershey (ed.), p. 455–468. The bacteriophage lambda. New York: Cold Spring Harbor Laboratory 1971.
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Horiuchi, T., Nagata, T. Mutations affecting growth of the Escherichia coli cell under a condition of DNA polymerase I-deficiency. Molec. Gen. Genet. 123, 89–110 (1973). https://doi.org/10.1007/BF00282992
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DOI: https://doi.org/10.1007/BF00282992