Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1976 Apr;18(1):235–244. doi: 10.1128/jvi.18.1.235-244.1976

Isolation and partial characterization of a corynebacteriophage beta, tox operator constitutive-like mutant lysogen of Corynebacterium diphtheriae.

J R Murphy, J Skiver, G McBride
PMCID: PMC515543  PMID: 815566

Abstract

We have isolated and partially characterized a beta-phage mutant lysogen of Corynebacterium diphtheriae, C7(betatoxct1+), which is partially insensitive to iron inhibition of diphtheria toxin production. tox expression by C7(betatoxct1+) was found to be partially constitutive. In the presence of concentrations of iron that almost completely inhibit the expression of diphtheria toxin by the wild type, C7(beta), the level of toxin production by C7(betatoxct1+) was found to be at least 25 times that of the parent. The purified tox gene product of C7(betatoxct1+) was immunologically and electrophoretically identical to, and equally as toxic as, diphtheria toxin purified from C7(beta). In addition, the partial N-terminal amino acid sequence was found to be identical to diphtheria toxin. This data strongly suggests that the mutation allowing for the constitutive expression of tox in C7(betatoxct1+) is outside of the structural gene. Furthermore, the constitutive expression of diphtheria toxin was found to be cis dominant in the double lysogen C7(betacrm45+/betatoxct1+). The data presented is consistent with the existence of a tox operator locus.

Full text

PDF
235

Images in this article

238Image
on p.238
240Image
on p.240
241Image
on p.241

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baseman J. B., Pappenheimer A. M., Jr, Gill D. M., Harper A. A. Action of diphtheria toxin in the guinea pig. J Exp Med. 1970 Dec 1;132(6):1138–1152. doi: 10.1084/jem.132.6.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Collier R. J. Diphtheria toxin: mode of action and structure. Bacteriol Rev. 1975 Mar;39(1):54–85. doi: 10.1128/br.39.1.54-85.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. ELEK S. D. The plate virulence test for diphtheria. J Clin Pathol. 1949 Nov;2(4):250–258. doi: 10.1136/jcp.2.4.250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. FREEMAN V. J. Studies on the virulence of bacteriophage-infected strains of Corynebacterium diphtheriae. J Bacteriol. 1951 Jun;61(6):675–688. doi: 10.1128/jb.61.6.675-688.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gill D. M., Pappenheimer A. M., Jr Structure-activity relationships in diphtheria toxin. J Biol Chem. 1971 Mar 10;246(5):1492–1495. [PubMed] [Google Scholar]
  6. Gill D. M., Uchida T., Singer R. A. Expression of diphtheria toxin genes carried by integrated and nonintegrated phage beta. Virology. 1972 Dec;50(3):664–668. doi: 10.1016/0042-6822(72)90420-5. [DOI] [PubMed] [Google Scholar]
  7. Gros C., Labouesse B. Study of the dansylation reaction of amino acids, peptides and proteins. Eur J Biochem. 1969 Feb;7(4):463–470. doi: 10.1111/j.1432-1033.1969.tb19632.x. [DOI] [PubMed] [Google Scholar]
  8. HATANO M. Effect of iron concentration in the medium on phage and toxin production in a lysogenic, virulent Corynebacterium diphtheriae. J Bacteriol. 1956 Jan;71(1):121–122. doi: 10.1128/jb.71.1.121-122.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hagen F. S., Young E. T. Preparative polyacrylamide gel electrophoresis of ribonucleic acid. Identification of multiple molecular species of bacteriophage T7 lysozyme messenger ribonucleic acid. Biochemistry. 1974 Jul 30;13(16):3394–3400. doi: 10.1021/bi00713a033. [DOI] [PubMed] [Google Scholar]
  10. Holmes R. K., Barksdale L. Genetic analysis of tox+ and tox- bacteriophages of Corynebacterium diphtheriae. J Virol. 1969 Jun;3(6):586–598. doi: 10.1128/jvi.3.6.586-598.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. JACOB F., MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318–356. doi: 10.1016/s0022-2836(61)80072-7. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Matsuda M., Barksdale L. Phage-directed synthesis of diphtherial toxin in non-toxinogenic Corynebacterium diphtheriae. Nature. 1966 May 28;210(5039):911–913. doi: 10.1038/210911a0. [DOI] [PubMed] [Google Scholar]
  14. Michel A., Zanen J., Monier C., Crispeels C., Dirkx J. Partial characterization of diphtheria toxin and its subunits. Biochim Biophys Acta. 1972 Feb 29;257(2):249–256. doi: 10.1016/0005-2795(72)90276-0. [DOI] [PubMed] [Google Scholar]
  15. Mosteller R. D., Yanofsky C. Transcription of the tryptophan operon in Escherichia coli: rifampicin as an inhibitor of initiation. J Mol Biol. 1970 Mar;48(3):525–531. doi: 10.1016/0022-2836(70)90064-1. [DOI] [PubMed] [Google Scholar]
  16. Murphy J. R., Pappenheimer A. M., Jr, de Borms S. T. Synthesis of diphtheria tox-gene products in Escherichia coli extracts. Proc Natl Acad Sci U S A. 1974 Jan;71(1):11–15. doi: 10.1073/pnas.71.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pappenheimer A. M., Jr, Uchida T., Harper A. A. An immunological study of the diphtheria toxin molecule. Immunochemistry. 1972 Sep;9(9):891–906. doi: 10.1016/0019-2791(72)90163-2. [DOI] [PubMed] [Google Scholar]
  18. Roth J. R., Antón D. N., Hartman P. E. Histidine regulatory mutants in Salmonella typhimurium. I. Isolation and general properties. J Mol Biol. 1966 Dec 28;22(2):305–323. doi: 10.1016/0022-2836(66)90134-3. [DOI] [PubMed] [Google Scholar]
  19. Singer R. A. Temperature-sensitive mutants of toxinogenic corynebacteriophage beta. I. Genetics. Virology. 1973 Oct;55(2):347–356. doi: 10.1016/0042-6822(73)90174-8. [DOI] [PubMed] [Google Scholar]
  20. Uchida T., Gill D. M., Pappenheimer A. M., Jr Mutation in the structural gene for diphtheria toxin carried by temperate phage . Nat New Biol. 1971 Sep 1;233(35):8–11. doi: 10.1038/newbio233008a0. [DOI] [PubMed] [Google Scholar]
  21. Uchida T., Pappenheimer A. M., Jr, Greany R. Diphtheria toxin and related proteins. I. Isolation and properties of mutant proteins serologically related to diphtheria toxin. J Biol Chem. 1973 Jun 10;248(11):3838–3844. [PubMed] [Google Scholar]
  22. Uchida T., Pappenheimer A. M., Jr, Harper A. A. Diphtheria toxin and related proteins. 3. Reconstitution of hybrid "diphtheria toxin" from nontoxic mutant proteins. J Biol Chem. 1973 Jun 10;248(11):3851–3854. [PubMed] [Google Scholar]
  23. Uchida T., Pappenheimer A. M., Jr, Harper A. A. Diphtheria toxin and related proteins. II. Kinetic studies on intoxication of HeLa cells by diphtheria toxin and related proteins. J Biol Chem. 1973 Jun 10;248(11):3845–3850. [PubMed] [Google Scholar]
  24. Woods K. R., Wang K. T. Separation of dansyl-amino acids by polyamide layer chromatography. Biochim Biophys Acta. 1967 Feb 21;133(2):369–370. doi: 10.1016/0005-2795(67)90078-5. [DOI] [PubMed] [Google Scholar]
  25. Wuesthoff G., Bauerle R. H. Mutations creating internal promoter elements in the tryptophan operon of Salmonella typhimurium. J Mol Biol. 1970 Apr 14;49(1):171–196. doi: 10.1016/0022-2836(70)90384-0. [DOI] [PubMed] [Google Scholar]
  26. YONEDA M., PAPPENHEIMER A. M., Jr Some effects of iron deficiency on the extracellular products released by toxigenic and nontoxigenic strains of Corynebacterium diphtheriae. J Bacteriol. 1957 Aug;74(2):256–264. doi: 10.1128/jb.74.2.256-264.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES