Structural features of DNA in competent Bacillus subtilis
Tóm tắt
For efficient transformation with B. subtilis, recipient cells must be grown to the state termed competence. Previous findings indicated that such competent cells contained DNA which exhibited about 5% single-strandedness. In this work, the physico-chemical properties of this DNA are compared to artificially “nicked” DNA. Evidence is presented that breakdown of the host DNA occurs during growth to competence. Inhibition of this breakdown also prevents the formation of partially single-stranded chromosomes within the competent cells. Use of this DNA as donor in transformation studies indicated a deficiency in biological activity within specific genes. Of three models considered, it is concluded that the results are best explained by the occurrence of single-stranded gaps within the chromosomes of competent cells.
Tài liệu tham khảo
Anagnostopoulos, C., Spizizen, J.: Requirements for the transformation in Bacillus subtilis. J. Bact. 81, 741–746 (1961).
Bode, V. C.: Single-strand scissions induced in circular and linear λ by the presence of dithiothreitol and other reducing agents. J. molec. Biol. 26, 125–129 (1967).
Bodmer, W. F.: Recombination and integration in Bacillus subtilis transformation: Involvement of DNA synthesis. J. molec. Biol. 14, 534–557 (1965).
Borenstein, S., Ephrati-Elizur, E.: Spontaneous release of DNA in sequential genetic order by Bacillus subtilis. J. molec. Biol. 45, 137–152 (1969).
Cahn, F. H., Fox, M. S.: Fractionation of transformable bacteria from competent cultures of B. subtilis on renograffin gradients. J. Bact. 95, 867–875 (1968).
Chen, K. C., Ravin, A. W.: Heterospecific transformation of Pneumococcus and Streptococcus 11: Dependence of relative efficiency of marker integration on host genome. J. molec. Biol. 22, 123–134 (1966).
Chilton, M. D.: Transforming activity in both complementary strands of Bacillus subtilis DNA. Science 157, 817–819 (1967).
Dubnau, D., Goldthwaite, C., Smith, I., Marmur, J.: Genetic mapping in Bacillus subtilis. J. molec. Biol. 27, 163–185 (1967).
Ephrati-Elizur, E.: Spontaneous transformation in Bacillus subtilis. Genet. Res. Camb. 11, 83–96 (1968).
Erickson, R. J., Braun, W.: Apparent dependence of transformation on the stage of deoxyribonucleic acid replication of recipient cells. Bact. Rev. 32, 291–296 (1968).
Flamm, W. G., Bond, H. E., Burr, H. E.: Density gradient centrifugation of DNA in a fixed-angle rotor—a higher order of resolution. Biochim. biophys. Acta (Amst.) 129, 310–319 (1966).
Guerrini, F., Fox, M. S.: Genetic heterozygosity in pneumococcal transformation. Proc. nat. Acad. Sci. (Wash.) 59, 429–436 (1968).
Hadden, C., Nester, E. W.: Purification of competent cells in the Bacillus subtilis transformation system. J. Bact. 95, 876–885 (1968).
Hadden, C., Nester, E. W.: DNA synthesis in the development of competence in Bacillus subtilis. Bact. Proc. p. 47 (1970).
Harris, W. J.: Structure and function of DNA during transformation. Ph. D. thesis, University of Dundee (1969).
Harris, W. J.: Communicated 8th Int. congress. Biochem. (1970).
—, Barr, G. C.: Some properties of DNA in competent Bacillus subtilis. J. molec. Biol. 39, 245–255 (1969).
Hoch, J. A., Barat, M., Anagnostopoulos, C.: Transformation and transduction in recombination-defective mutants of Bacillus subtilis. J. Bact. 93, 1925–1937 (1967).
Kelly, M. S., Pritchard, R. H.: Unstable linkage between genetic markers in transformation. J. Bact. 89, 1314–1321 (1965).
Kirby, K. S.: A new method for the isolation of DNA: evidence on the nature of bonds between deoxyribonucleic acid and protein. Biochem. J. 66, 495–503 (1957).
—: Preparation of some deoxyribonucleic acid-protein complexes from rat liver homogenates. Biochem. J. 70, 260–265 (1958).
Lacks, S.: Integration efficiency and genetic recombination in pneumococcal transformation. Genetics 53, 207–235 (1966).
Laird, C. D., Wang, L., Bodmer, W. F.: Recombination of DNA replication in Bacillus subtilis transformation. Mutation Res. 6, 205–209 (1968).
Mahler, I., Cahoon, M., Marmur, J.: Bacillus subtilis deoxyribonucleic acid transfer in PBS2 transduction. J. Bact. 87, 1423–1428 (1964).
McCarthy, C., Nester, E. W.: Macromolecular synthesis in newly transformed cells of Bacillus subtilis. J. Bact. 94, 131–140 (1967).
Ottolenghi, E., Hotchkiss, R. D.: Release of genetic transforming agent from pneumococcal cultures during growth and disintegration. J. exp. Med. 116, 491–519 (1962).
Rudner, R., Karkas, J. D., Chargaff, E.: Separation of Bacillus subtilis DNA into complementary strands 1: Biological properties. Proc. nat. Acad. Sci. (Wash.) 60, 630–635 (1968).
Rupp, W. D., Howard-Flanders, P.: Discontinuities in the DNA synthesised in an excision-defective strain of Escherichia coli following ultraviolet irradiation. J. molec. Biol. 31, 291–304 (1968).
Sicard, A. M., Ephrussi-Taylor, H.: Genetic recombination in DNA-induced transformation of pneumococcus 11: mapping of the amiA region. Genetics 52, 1207–1227 (1965).
Singh, R. N., Pitale, M. P.: Enrichment of Bacillus subtilis transformants by zonal centrifugation. Nature (Lond.) 213, 1262–1263 (1967).
Steinhart, W. L., Herriott, R. M.: Fate of recipient deoxyribonucleic acid during transformation in Haemophilus influenzae. J. Bact. 96, 1718–1724 (1968).
Tevethia, M. J., Mandel, M.: Nature of ethylenediamino-tetra acetic acid requirement for transformation of Bacillus subtilis with single-stranded deoxyribonucleic acid. J. Bact. 101, 844–850 (1970).
Tomasz, A.: The competent state in genetic transformation. Ann. Rev. Genet. 3, 217–232 (1969).
Vinograd, J., Hearst, J. E.: Equilibrium sedimentation of macromolecules and viruses in a density gradient. Fortschr. Chem. org. Naturstoffe 10, 373–410 (1962).
—, Morris, J., Davidson, N., Dove, W. F.: The bouyant behaviour of viral and bacterial DNA in alkaline CsCl. Proc. nat. Acad. Sci. (Wash.) 49, 12–17 (1963).
—, Lebowitz, J., Watson, R.: Early and late helix-coil transitions in closed circular DNA. The number of superhelical turns in Polyoma DNA. J. molec. Biol. 33, 173–197 (1968).
Weiss, B., Richardson, C. C.: Enzymatic breakage and joining of DNA 1: Repair of single-strand breaks in DNA by an enzyme system from Escherichia coli infected with T4 bacteriophage. Proc. nat. Acad. Sci. (Wash.) 57, 1021–1028 (1967).
Yoshikawa, H.: Mutations resulting from the transformation of Bacillus subtilis. Genetics 54, 1201–1214 (1966).