Summary
The action of nitrous acid on H. influenzae transforming DNA has been studied. Two of the antibiotic resistance markers studied, marker S (streptomycin) and marker N (novobiocin) show a difference in sensitivity towards nitrous acid. For these two markers, the loss of transforming activity is in accordance with a theory of transforming DNA proposed by Bresler and coll. In this theory the inactivation is related to the existence of damages that reduce the length of the donor DNA that is integrated. For the third marker studied, a K marker (kanamycin) the shape of the inactivation curve is somewhat different and shows the existence of an exponential component. The inactivation of the streptomycin-novobiocin linkage group (weakly linked markers) is additive, whereas the inactivation of the streptomycine-kanamycine group (closely linked markers) shows an overlapping effect which can be accounted for by the theory. These results are discussed in the text.
Furthermore, the kinetic of induction by nitrous acid of “reversible” DNA molecules has been studied. This has been done by means of measurements of the transforming activity of nitrous acid treated DNA before and after heat denaturation and by hydroxylapatite chromatography of the heated DNA. It is shown that the kinetic of induction of “reversibility” is the same for the three unlinked markers studied. The direct estimation of the size of the crosslinked fraction indicates that the crosslinking effect of nitrous acid is not a major factor in marker inactivation.
Finally, in view of our own results and of those obtained by others, we discuss the failure of nitrous acid to be mutagenic on native H. influenzae DNA. This failure, which contrasts with the result obtained on B. subtilis and Pneumococcal DNA, cannot be attributed to the existence of the nitrous induced crosslinks.
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Références
Alberts, B. M.: Ph. D. Dissertation, Université de Harward (1965).
Bach, M. L.: Etude de l'action de divers agents sur l'ADN transformant d'Hemophilus influenzae. Thèse, Université de Strasbourg, 1968.
—, Luzzati, D., Chevallier, M. R.: Action du peroxyde succinique sur l'activité transformante de l'acide deoxyribonucléique de l'Hemophilus influenzae. Biochem. biophys. Acta (Amst.) 25, 76 (1963).
Becker, E. J. J., Zimmerman, B. K., Geiduscheck, E. P.: Structure and function of crosslinked DNA. I. Reversible denaturation and Bacillus subtilis transformation. J. molec. Biol. 8, 377–391, (1964).
Bernardi, G.: Chromatography of nucleic acids on hydroxyapatite. Nature (Lond.) 206, 779–783 (1965).
Bresler, S. E., Kalinin, V. M., Perumov, D. A.: Inactivation and mutagenesis on isolated DNA. I. Theory of inactivation of transforming DNA. Mutation Res. 4, 389–398 (1967).
—: Inactivation and mutagenesis on isolated DNA. II. Kinetics of mutagenesis and efficiency of different mutagens. Mutation Res. 5, 1–14 (1968).
—: Inactivation and mutagenesis on isolated DNA. IVI. Additivity of action of different agents on transforming DNA. Mutation Res. 5, 209–215 (1968).
—: Inactivation and mutagenesis on isolated DNA. IV. Possibility of integration of lethal damage into chromosome of B. Subtilis during transformation. Mutation Res. 5, 329–341 (1968).
Cabrera-Juarez, E., Herriott, R. M.: Ultraviolet inactivation of native and denatured transforming DNA from Haemophilus influenzae. J. Bact. 85, 671–675 (1963).
Canner, M. K., Spizizen, J.: Inactivation of Bacillus Subtilis DNA by mutagenic agents. J. Bact. 89, 915–916 (1965).
Chevallier, M. R.: resultats non publiés (1969).
—, Bernardi, G.: Transformation by heat-denatured deoxyribonucleic acid. J. molec. Biol. 11, 658, (1965).
—: Residual transforming activity of denatured Haemophilus influenzae DNA. J. molec. Biol. 32, 437–452 (1968).
—, Londos-Gagliardi, D., Nicolaïeff, A., Aubel-Sadron, G.: Halogénation des acides nucléiques. VI. Influence de la bromuration sur l'activité transformante du DNA d'Hemophilus influenzae. Bull. Soc. Chim. biol. (Paris) 50, 393–407 (1968).
Geiduscheck, E. P.: On the factors controlling the reversibility of DNA denaturation. J. molec. Biol. 4, 467–487 (1962).
— “Reversible” DNA. Proc. nat. Acad. Sci. (Wash.) 47, 951–955 (1961).
Goodgal, S. H., Herriott, R. M.: Studies of transformation of Hemophilus influenzae. I. Competence. J. gen. Physiol. 44, 1201–1227 (1961).
—, Horn-Postel, E.: Nitrous acid mutation of transforming DNA: Consideration of mode of action. Science 148, 1095–1097 (1965).
Haldane, J. B. S.: The combination of linkage values and the calculation of distances between loci of linked factors. J. Genet. 8, 299–311 (1919).
Horn, E., Herriott, R. M.: The mutagenic action of nitrous acid on “single-stranded” (Denatured) Hemophilus Transforming DNA. Proc. nat. Acad. Sci. (Wash.) 48, 1409–1416 (1962)
Lacks, S.: Integration efficiency and genetic recombination in pneumococcal transformation. Genetics 53, 207–235 (1966).
—: Theoretical relationship between probality of Marker integration and length of donor DNA in pneumococcal transformation. J. molec. Biol. 37, 179–180 (1968).
Litman, R.: Genetic and chemical alterations in the transforming DNA of Pneumoccus caused by ultraviolet light and by nitrous acid. J. Chim. Phys. 58, 997–1007 (1961).
Luzzati, D.: The action of nitrous acid on transforming desoxyribonucleic acids. Biochem. biophys. Res. Commun. 9, 508–516 (1962).
—, Schweitz, H., Bach, M. L., Chevallier, M. R.: Action du peroxyde succinique sur l'ADN. J. Chim. Phys. 58, 1021–1027 (1961).
Iyer, V. N., Szybalski, W.: A molecular mechanism of mitomycin action: linking of complementary DNA strands. Proc. nat. Acad. Sci. (Wash.) 50, 355 (1963).
Michalka, J., Goodgal, S. H.: A genetic and physical map of the chromosome of H. influenzae. Sous presse (1969).
Nicolaïeff, A., Chevallier, M. R.: En préparation.
Postel, E. H., Goodgal, S. H.: Induction of mutations by nitrous acid on denatured Haemophilus influenzae. Deoxyribonucleic acid assayed directly by single-stranded transformation. J. Bact. 94, 1802–1803 (1967).
Rupert, C., Goodgal, S. H.: Shape of ultraviolet inactivation curves of transforming deoxyribonucleic acid. Nature (Lond.) 185, 556–557 (1960).
Schuster, H.: Die Reaktionsweise der Desoxyribonucleinsäure mit salpetriger Säure. Z. Naturforsch. 15b, 298–304 (1960).
Setlow, J. K., Brown, D. C., Boling, M. E., Mattingly, A., Gordon, M. P.: Repair of deoxyribonucleic acid in Haemophilus influenzae. I. X Ray sensitivity of ultravioletsensitive mutants and their behavior as hosts to ultraviolet-irradiated bacteriophage and transforming deoxyribonucleic acid. J. Bact. 95, 546–558 (1968).
Sluis, C. A. van, Stuy, J. H.: On the inactivation of transforming deoxyribonucleic acid by heat. Biochem. biophys. res. Commun. 7, 212–219 (1962).
Spencer, H. I., Herriott, R. M.: Development of competence of H. influenzae. J. Bact. 90, 911–920 (1965).
Stuy, J. H.: Inactivation of transforming deoxyribonucleic acid by nitrous acid. Biochem. biophys. Res. Commun. 6, 328–332 (1961).
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Chevallier, M.R., Greth, M.L. Etude de l'action de l'acide nitreux sur le DNA transformant d'Haemophilus influenzae . Molec. Gen. Genet. 105, 344–360 (1969). https://doi.org/10.1007/BF00277589
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DOI: https://doi.org/10.1007/BF00277589