Abstract
Diphtherial toxin is produced in maximal yields by Corynebacterium diphtheriae (C7(beta tox+) only when iron is present in growth-limiting amounts. Toxin production is markedly decreased under high-iron conditions. We studied the role of the bacteriophage beta genome in this apparent regulation of toxin production by iron. Using a passive immune hemolysis assay to detect toxin antigen production in individual plaques, we identified rare phage mutants that were toxinogenic in high-iron medium. Lysogenic derivatives of C. diphtheriae C7 harboring such phage mutants were constructed. The lysogens were compared with wild-type strain C7(beta) for their ability to produce toxin in deferrated liquid medium containing varying amounts of added iron. Quantitative tests for extracellular toxin were performed by competitive-binding radioimmunoassays. We identified phenotypically distinct mutant strains that produced slightly, moderately, or greatly increased yields of toxin antigen under high-iron conditions. The toxin produced by the mutant lysogens was biologically active and immunochemically indistinguishable from wild-type toxin. Complementation experiments demonstrated that the phage mutation designated tox-201 had a cis-dominant effect on the expression of the toxin structural gene of phage beta. The characteristics of the tox-201 mutation suggest that it defines a regulatory locus of phage beta that is involved in control of toxinogenesis by iron in C. diphtheriae.
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Arditti R., Grodzicker T., Beckwith J. Cyclic adenosine monophosphate-independent mutants of the lactose operon of Escherichia coli. J Bacteriol. 1973 May;114(2):652–655. doi: 10.1128/jb.114.2.652-655.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BARDSDALE W. L., PAPPENHEIMER A. M., Jr Phage-host relationships in nontoxigenic and toxigenic diphtheria bacilli. J Bacteriol. 1954 Feb;67(2):220–232. doi: 10.1128/jb.67.2.220-232.1954. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BAUTZ-FREESE E., FREESE E. Induction of reverse mutations and cross reactivation of nitrous acid-treated phage T4. Virology. 1961 Jan;13:19–30. doi: 10.1016/0042-6822(61)90027-7. [DOI] [PubMed] [Google Scholar]
- Bacha P., Murphy J. R. Isolation and characterization of extragenic suppressor strains of Corynebacterium diphtheriae. J Bacteriol. 1978 Dec;136(3):1135–1142. doi: 10.1128/jb.136.3.1135-1142.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barksdale L. Corynebacterium diphtheriae and its relatives. Bacteriol Rev. 1970 Dec;34(4):378–422. doi: 10.1128/br.34.4.378-422.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barksdale L. I. : Lysogenic Conversions in Bacteria. Bacteriol Rev. 1959 Dec;23(4):202–212. doi: 10.1128/br.23.4.202-212.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Cryz S. J., Welkos S. L., Holmes R. K. Immunochemical studies of diphtherial toxin and related nontoxic mutant proteins. Infect Immun. 1980 Dec;30(3):835–846. doi: 10.1128/iai.30.3.835-846.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Crombrugghe B., Chen B., Gottesman M., Pastan I., Varmus H. E., Emmer M., Perlman R. L. Regulation of lac mRNA synthesis in a soluble cell-free system. Nat New Biol. 1971 Mar 10;230(10):37–40. doi: 10.1038/newbio230037a0. [DOI] [PubMed] [Google Scholar]
- EDWARDS D. C., SEAMER P. A. The uptake of iron by Corynebacterium diphtheriae growing in submerged culture. J Gen Microbiol. 1960 Jun;22:705–712. doi: 10.1099/00221287-22-3-705. [DOI] [PubMed] [Google Scholar]
- FREEMAN V. J., MORSE I. U. Further observations on the change to virulence of bacteriophage-infected a virulent strains of Corynebacterium diphtheria. J Bacteriol. 1952 Mar;63(3):407–414. doi: 10.1128/jb.63.3.407-414.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- GROMAN N. B., BOBB D. The inhibition of adsorption of Corynebacterium diphtheriae phage by tween 80. Virology. 1955 Sep;1(3):313–323. doi: 10.1016/0042-6822(55)90027-1. [DOI] [PubMed] [Google Scholar]
- GROMAN N. B. Conversion in Corynebacterium diphtheriae with phages originating from nontoxigenic strains. Virology. 1956 Dec;2(6):843–844. doi: 10.1016/0042-6822(56)90066-6. [DOI] [PubMed] [Google Scholar]
- GROMAN N. B. Evidence for the induced nature of the change from nontoxigenicity to toxigenicity in Corynebacterium diphtheriae as a result of exposure to specific bacteriophage. J Bacteriol. 1953 Aug;66(2):184–191. doi: 10.1128/jb.66.2.184-191.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Griffiths E., Humphreys J., Leach A., Scanlon L. Alterations in the tRNA's of Escherichia coli recovered from lethally infected animals. Infect Immun. 1978 Nov;22(2):312–317. doi: 10.1128/iai.22.2.312-317.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Holmes R. K. Characterization and genetic mapping of nontoxinogenic (tox) mutants of corynebacteriophage beta. J Virol. 1976 Jul;19(1):195–207. doi: 10.1128/jvi.19.1.195-207.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Holmes R. K., Perlow R. B. Quantitative assay of diphtherial toxin and of immunologically cross-reacting proteins by reversed passive hemagglutination. Infect Immun. 1975 Dec;12(6):1392–1400. doi: 10.1128/iai.12.6.1392-1400.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Johnston H. M., Barnes W. M., Chumley F. G., Bossi L., Roth J. R. Model for regulation of the histidine operon of Salmonella. Proc Natl Acad Sci U S A. 1980 Jan;77(1):508–512. doi: 10.1073/pnas.77.1.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanei C., Uchida T., Yoneda M. Isolation from corynebacterium diphtheriae C7(beta) of bacterial mutants that produce toxin in medium with excess iron. Infect Immun. 1977 Oct;18(1):203–209. doi: 10.1128/iai.18.1.203-209.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keller E. B., Calvo J. M. Alternative secondary structures of leader RNAs and the regulation of the trp, phe, his, thr, and leu operons. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6186–6190. doi: 10.1073/pnas.76.12.6186. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laird W., Groman N. Isolation and characterization of tox mutants of corynebacteriophage beta. J Virol. 1976 Jul;19(1):220–227. doi: 10.1128/jvi.19.1.220-227.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laird W., Groman N. Orientation of the tox gene in the prophage of corynebacteriophage beta. J Virol. 1976 Jul;19(1):228–231. doi: 10.1128/jvi.19.1.228-231.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lightfoot H. N., Iglewski B. H. Synthesis of diphtheria toxin in E. coli cell-free lysate. Biochem Biophys Res Commun. 1974 Jan 23;56(2):351–357. doi: 10.1016/0006-291x(74)90849-3. [DOI] [PubMed] [Google Scholar]
- Matsuda M., Barksdale L. System for the investigation of the bacteriophage-directed synthesis of diphtherial toxin. J Bacteriol. 1967 Feb;93(2):722–730. doi: 10.1128/jb.93.2.722-730.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsuda M., Kanei C., Yoneda M. A phage-mutant directed synthesis of a fragment of diptheria toxin protein. Biochem Biophys Res Commun. 1972 Jan 14;46(1):43–49. doi: 10.1016/0006-291x(72)90627-4. [DOI] [PubMed] [Google Scholar]
- Matsuda M., Kanei C., Yoneda M. Degree of expression of the tox + gene introduced into corynebacteria by phages derived from diphtheria bacilli having different capacities to produce toxin. Biken J. 1971 Sep;14(3):365–368. [PubMed] [Google Scholar]
- Murphy J. R., Michel J. L., Teng M. Evidence that the regulation of diphtheria toxin production is directed at the level of transcription. J Bacteriol. 1978 Aug;135(2):511–516. doi: 10.1128/jb.135.2.511-516.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Murphy J. R., Skiver J., McBride G. Isolation and partial characterization of a corynebacteriophage beta, tox operator constitutive-like mutant lysogen of Corynebacterium diphtheriae. J Virol. 1976 Apr;18(1):235–244. doi: 10.1128/jvi.18.1.235-244.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- PARSONS E. I. Induction of toxigenicity in non-toxigenic strains of C. diphtheriae with bacteriophages derived from non-toxigenic strains. Proc Soc Exp Biol Med. 1955 Oct;90(1):91–93. doi: 10.3181/00379727-90-21948. [DOI] [PubMed] [Google Scholar]
- Pappenheimer A. M., Jr Diphtheria toxin. Annu Rev Biochem. 1977;46:69–94. doi: 10.1146/annurev.bi.46.070177.000441. [DOI] [PubMed] [Google Scholar]
- RAYNAUD M., TURPIN A., MANGALO R., BIZZINI B. Croissance et toxinogenèse. Ann Inst Pasteur (Paris) 1954 Dec;87(6):599–616. [PubMed] [Google Scholar]
- Righelato R. C., van Hemert P. A. Growth and toxin synthesis in batch and chemostat cultures of Corynebacterium diphtheriae. J Gen Microbiol. 1969 Nov;58(3):403–410. doi: 10.1099/00221287-58-3-403. [DOI] [PubMed] [Google Scholar]
- Silverstone A. E., Arditti R. R., Magasanik B. Catabolite-insensitive revertants of lac promoter mutants. Proc Natl Acad Sci U S A. 1970 Jul;66(3):773–779. doi: 10.1073/pnas.66.3.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Smith W. P. Cotranslational secretion of diphtheria toxin and alkaline phosphatase in vitro: involvement of membrane protein(s). J Bacteriol. 1980 Mar;141(3):1142–1147. doi: 10.1128/jb.141.3.1142-1147.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith W. P., Tai P. C., Murphy J. R., Davis B. D. Precursor in cotranslational secretion of diphtheria toxin. J Bacteriol. 1980 Jan;141(1):184–189. doi: 10.1128/jb.141.1.184-189.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stauffer G. V., Zurawski G., Yanofsky C. Single base-pair alterations in the Escherichia coli trp operon leader region that relieve transcription termination at the trp attenuator. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4833–4837. doi: 10.1073/pnas.75.10.4833. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Uchida T., Kanei C., Yoneda M. Mutations of corynephage beta that affect the yield of diphtheria toxin. Virology. 1977 Apr;77(2):876–879. doi: 10.1016/0042-6822(77)90511-6. [DOI] [PubMed] [Google Scholar]
- 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]
- Uchida T., Pappenheimer A. M., Jr, Harper A. A. Reconstitution of diphtheria toxin from two nontoxic cross-reacting mutant proteins. Science. 1972 Feb 25;175(4024):901–903. doi: 10.1126/science.175.4024.901. [DOI] [PubMed] [Google Scholar]
- Uchida T., Yoneda M. Estimation of the time required for the process of diphtheria toxin formation. Biken J. 1967 Sep;10(3):121–128. [PubMed] [Google Scholar]
- Uchida T., Yoneda M. Evidence for the association of membrane with the site of toxin synthesis in Corynebacterium diphtheriae. Biochim Biophys Acta. 1967 Aug 22;145(1):210–213. doi: 10.1016/0005-2787(67)90681-8. [DOI] [PubMed] [Google Scholar]
- Welkos S. L., Holmes R. K. Characterization of a screening test for diphtherial toxin antigen produced by individual plaques of corynebacteriophages. J Clin Microbiol. 1979 Jun;9(6):693–698. doi: 10.1128/jcm.9.6.693-698.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Welkos S. L., Holmes R. K. Regulation of toxinogenesis in Corynebacterium diphtheriae. II. Genetic mapping of a tox regulatory mutation in bacteriophage beta. J Virol. 1981 Mar;37(3):946–954. doi: 10.1128/jvi.37.3.946-954.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]