Skip to main content
Log in

Streptomycin-resistant, asporogenous mutant of Bacillus subtilis

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A streptomycin-resistant mutant of Bacillus subtilis that is also asporogenous, was isolated and partially characterized. This strain, SRB15, sporulated at a frequency of about 1% compared to the wild type frequency of greater than 70%. The two phenotypes were inseparable by transformation, suggesting that this strain carries a single mutation that causes it to be both streptomycin-resistant and spore-minus. The mutation cotransduces with cysA, the closest auxotrophic marker to the “ribosomal region” of the B. subtilis chromosome, with a frequency of 68%. SRB15 showed no cross resistance to other antibioties tested, including the aminoglycosides kanamycin, neomycin and spectinomycin. Ribosomes obtained from the mutant were at least 200-fold more resistant in vitro to streptomycin than were wild type ribosomes in the translation of phage SPO1 RNA. The kinetics of in vitro translation of this natural message were indistinguishable for mutant and wild type ribosomes. The level of misreading, as measured by poly(U)-directed isoleucine incorporation, by mutant ribosomes was less than that by wild type ribosomes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Anagnostopoulous, C., Spizizen, J.: Requirement for transformation in Bacillus subtilis. J. Bact. 80, 741–746 (1961)

    Google Scholar 

  • Arceneaux, J., Sueoka, N.: Two species of Bacillus subtilis tyrosine transfer ribonucleic acid. J. biol. Chem. 244, 5959–5966 (1966)

    Google Scholar 

  • Bott, K., Graham, S., Chambliss, G.: Translational control and its relevance to sporulation. Colloques internationaux CNRS-Regulation de la sporulation microbienne 227, 96–107 (1973)

    Google Scholar 

  • Bott, K., Wilson, G.: Development of competence in the Bacillus subtilis transformation system. J. Bact. 94, 562–570 (1967)

    Google Scholar 

  • Boylan, R., Mendelson, N., Brooks, D., Young, F.: Regulation of the bacterial cell wall: Analysis of a mutant of Bacillus subtilis defective in biosynthesis of teichoic acid. J. Bact. 121, 670–674 (1972)

    Google Scholar 

  • Cannon, J., Graham, S., Bott, K.: Altered sporulation among spectinomycin resistant mutants of Bacillus subtilis. In: Microbiology — 1976 (ed. D. Schlessinger), pp. 220–227. Washington, D.C.: American Society for Microbiology 1976

    Google Scholar 

  • Chakrabarti, S., Gorini, L.: Growth of bacteriophages MS2 and T7 on streptomycin-resistant mutants of Escherichia coli. J. Bact. 121, 670–674 (1975)

    Google Scholar 

  • Chambliss, G., Legault-Demare, L.: Template discrimination by the initiation factor fraction from the ribosomes of sporulating Bacillus subtilis. In: Spores VI (ed. P. Gerhardt, R. Costilow, and H. Sadoff), pp. 314–317. Washington, D.C.: American Society for Microbiology 1975

    Google Scholar 

  • Davies, J., Gilbert, W., Gorini, L.: Streptomycin, suppression and the code. Proc. nat. Acad. Sci. (Wash.) 51, 883–890 (1964)

    Google Scholar 

  • Domoto, T., Kobayashi, K., Kobayashi, Y.: Erythromycin-resistant, conditional asporogenous mutant of Bacillus subtilis. In: Spores VI (ed. P. Gerhardt, R. Costilow, and H. Sadoff), pp. 307–313. Washington, D.C.: American Society for Microbiology 1975

    Google Scholar 

  • Dubnau, D., Goldwaithe, C., Smith, I., Marmur, J.: Genetic mapping in Bacillus subtilis. J. molec. Biol. 27, 163–185 (1967)

    Google Scholar 

  • Fortnagel, P., Berman, R., Hafeman, B., Lengelsen, C.: Structural and functional alterations of Bacillus subtilis ribosomes. In: Spores VI (ed. P. Gerhardt, R. Costilow, and H. Sadoff), pp. 301–306. Washington, D.C.: American Society for Microbiology 1975

    Google Scholar 

  • Galas, D., Branscomb, E.: Ribosome slowed by mutation to streptomycin resistance. Nature (Lond.) 262, 617–619 (1976)

    Google Scholar 

  • Goldberg, M., Steitz, J.: Cistron specificity of 30S ribosomes heterologously reconstituted with components from Escherichia coli and Bacillus stearothermophilus. Biochemistry 13, 2123–2129 (1974)

    Google Scholar 

  • Graham, S., Bott, K.: Antibiotic-resistant mutants of Bacillus subtilis conditional for sporulation. Molec. gen. Genet. 137, 227–237 (1975)

    Google Scholar 

  • Guha, S.: Ribosomal proteins of Bacillus subtilis vegetative and sporulating cells. Molec. gen. Genet. 138, 299–307 (1975)

    Google Scholar 

  • Harford, N., Sueoka, N.: Chromosomal location of antibiotic resistance markers in Bacillus subtilis. J. molec. Biol. 51, 267–286 (1970)

    Google Scholar 

  • Held, W., Gette, W., Nomura, M.: Role of 16S ribosomal ribonucleic acid and the 30S ribosomal protein S12 in the initiation of natural messenger ribonucleic acid translation. Biochemistry 13, 2115–2122 (1974)

    Google Scholar 

  • Idriss, J., Halvorson, H.: The nature of ribosomes of spores of Bacillus cereus T and Bacillus megaterium. Arch. Biochem. Biophys. 133, 442–453 (1969)

    Google Scholar 

  • Jamet, C., Anagnostopoulos, C.: Etude d'une mutation tres faiblement transformable au locus de la threonine desaminase de Bacillus subtilis. Molec. gen. Genet. 105, 225–242 (1969)

    Google Scholar 

  • Kaneko, I., Doi, R.: Alteration of valyl-sRNA during sporulation of Bacillus subtilis. Proc. nat. Acad. Sci (Wash.) 55, 564–571 (1966)

    Google Scholar 

  • Kobayashi, Y.: Activation of dormant spore ribosomes during germination. II. Existence of defective ribosomal subunits in dormant spore ribosomes. In: Spores V (ed. H. Halvorson, R. Hanson, and L. Campbell), pp. 269–276. Washington, D.C.: American Society for Microbiology 1972

    Google Scholar 

  • Lazzarini, R.: Differences in lysine-sRNA from spores and vegetative cells of Bacillus subtilis. Proc. nat. Acad. Sci. (Wash.) 56, 185–190 (1966)

    Google Scholar 

  • Legault-Demare, L., Chambliss, G.: Natural messenger ribonucleic acid directed cell-free protein-synthesizing system of Bacillus subtilis. J. Bact. 120, 1300–1307 (1974)

    Google Scholar 

  • Legault-Demare, L., Chambliss, G.: Selective messenger translation by Bacillus subtilis ribosomes. Molec. gen. Genet. 142, 277–287 (1975)

    Google Scholar 

  • Leighton, T.: Sporulation-specific translational discrimination in Bacillus subtilis. J. molec. Biol. 86, 855–863 (1974)

    Google Scholar 

  • Lowry, O., Rosebrough, N., Farr, A., Randall, R.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • Northrup, J., Slepecky, R.: Sporulation mutants induced by heat in Bacillus subtilis. Science 155, 838–839 (1967)

    Google Scholar 

  • Ozaki, M., Mizushima, S., Nomura, M.: Identification and functional characterization of the protein controlled by the streptomycin-resistant locus in Escherichia coli. Nature (Lond.) 222, 333–339 (1969)

    Google Scholar 

  • Rhaese, H., Grade, R., Dichtelmuller, H.: Studies on the control of development: Correlation of initiation of differentiation with synthesis of highly phosphorylated nucleotides in Bacillus subtilis. Europ. J. Biochem. 64, 205–213 (1976)

    Google Scholar 

  • Schaeffer, P., Millet, J., Aubert, J.: Catabolic repression of bacterial sporulation. Proc. nat. Acad. Sci. (Wash.) 54, 704–711 (1965)

    Google Scholar 

  • Singhal, R., Vold, B.: Changes in transfer ribonucleic acids of Bacillus subtilis during different growth phases. Nucleic Acid Res. 3, 1249–1260 (1976)

    Google Scholar 

  • Sparling, P., Modolell, J., Takeda, Y., Davis, B.: Ribosomes from Escherichia coli merodiploids heterozygous for resistance to streptomycin and spectinomycin. J. molec. Biol. 37, 402–412 (1968)

    Google Scholar 

  • Staal, S., Hoch, J.: Conditional dihydrostreptomycin resistance in Bacillus subtilis. J. Bact. 110, 202–207 (1972)

    Google Scholar 

  • Steinberg, W.: Sporulation of Bacillus subtilis cells: In vivo evidence for phenotypic modification of a thermosensitive lysyltransfer ribonucleic acid synthetase. In: Spores VI (ed. P. Gerhardt, R. Costilow, and H. Sadoff), pp. 290–300. Washington, D.C.: American Society for Microbiology 1975

    Google Scholar 

  • Tipper, D., Johnson, C., Leighton, T.: Bacillus subtilis ribosomal mutants unable to sporulate at 47 C. In: Abstracts of the Annual Meeting of the American Society for Microbiology 1976

  • Woese, C.: Unusual ribosomal protein occurring during spore germination. J. Bact. 82, 695–701 (1961)

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by F. Gros

Rights and permissions

Reprints and permissions

About this article

Cite this article

Campbell, K.M., Chambliss, G.H. Streptomycin-resistant, asporogenous mutant of Bacillus subtilis . Molec. Gen. Genet. 158, 193–200 (1977). https://doi.org/10.1007/BF00268313

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00268313

Keywords

Navigation