Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1988 May;56(5):1066–1069. doi: 10.1128/iai.56.5.1066-1069.1988

Inhibitors of receptor-mediated endocytosis block the entry of Bacillus anthracis adenylate cyclase toxin but not that of Bordetella pertussis adenylate cyclase toxin.

V M Gordon 1, S H Leppla 1, E L Hewlett 1
PMCID: PMC259763  PMID: 2895741

Abstract

Bordetella pertussis and Bacillus anthracis produce extracytoplasmic adenylate cyclase toxins (AC toxins) with shared features including activation by calmodulin and the ability to enter target cells and catalyze intracellular cyclic AMP (cAMP) production from host ATP. The two AC toxins were evaluated for sensitivities to a series of inhibitors of known uptake mechanisms. Cytochalasin D, an inhibitor of microfilament function, abrogated the cAMP response to B. anthracis AC toxin (93%) but not the cAMP response elicited by B. pertussis AC toxin. B. anthracis-mediated intoxication of CHO cells was completely inhibited by ammonium chloride (30 mM) and chloroquine (0.1 mM), whereas the cAMP accumulation produced by B. pertussis AC toxin remained unchanged. The block of target cell intoxication by cytochalasin D could be bypassed when cells were first treated with anthrax AC toxin and then exposed to an acidic medium. These data indicate that despite enzymatic similarities, these two AC toxins intoxicate target cells by different mechanisms, with anthrax AC toxin entering by means of receptor-mediated endocytosis into acidic compartments and B. pertussis AC toxin using a separate, and as yet undefined, mechanism.

Full text

PDF
1066

Selected References

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

  1. Confer D. L., Slungaard A. S., Graf E., Panter S. S., Eaton J. W. Bordetella adenylate cyclase toxin: entry of bacterial adenylate cyclase into mammalian cells. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1984;17:183–187. [PubMed] [Google Scholar]
  2. Didsbury J. R., Moehring J. M., Moehring T. J. Binding and uptake of diphtheria toxin by toxin-resistant Chinese hamster ovary and mouse cells. Mol Cell Biol. 1983 Jul;3(7):1283–1294. doi: 10.1128/mcb.3.7.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Draper R. K., Simon M. I. The entry of diphtheria toxin into the mammalian cell cytoplasm: evidence for lysosomal involvement. J Cell Biol. 1980 Dec;87(3 Pt 1):849–854. doi: 10.1083/jcb.87.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Farfel Z., Friedman E., Hanski E. The invasive adenylate cyclase of Bordetella pertussis. Intracellular localization and kinetics of penetration into various cells. Biochem J. 1987 Apr 1;243(1):153–158. doi: 10.1042/bj2430153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Friedlander A. M. Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem. 1986 Jun 5;261(16):7123–7126. [PubMed] [Google Scholar]
  6. Hanski E., Farfel Z. Bordetella pertussis invasive adenylate cyclase. Partial resolution and properties of its cellular penetration. J Biol Chem. 1985 May 10;260(9):5526–5532. [PubMed] [Google Scholar]
  7. Hewlett E. L., Sauer K. T., Myers G. A., Cowell J. L., Guerrant R. L. Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin. Infect Immun. 1983 Jun;40(3):1198–1203. doi: 10.1128/iai.40.3.1198-1203.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hewlett E. L., Urban M. A., Manclark C. R., Wolff J. Extracytoplasmic adenylate cyclase of Bordetella pertussis. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1926–1930. doi: 10.1073/pnas.73.6.1926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hewlett E. L., Weiss A. A., Crane J. K., Pearson R. D., Anderson H. J., Myers G. A., Evans W. S., Hantske L. L., Kay H. D., Cronin M. J. Bordetella extracytoplasmic adenylate cyclase: actions as a bacterial toxin. Dev Biol Stand. 1985;61:21–26. [PubMed] [Google Scholar]
  10. Iglewski B. H., Kabat D. NAD-dependent inhibition of protein synthesis by Pseudomonas aeruginosa toxin,. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2284–2288. doi: 10.1073/pnas.72.6.2284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Leppla S. H. Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc Natl Acad Sci U S A. 1982 May;79(10):3162–3166. doi: 10.1073/pnas.79.10.3162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leppla S. H. Bacillus anthracis calmodulin-dependent adenylate cyclase: chemical and enzymatic properties and interactions with eucaryotic cells. Adv Cyclic Nucleotide Protein Phosphorylation Res. 1984;17:189–198. [PubMed] [Google Scholar]
  14. Middlebrook J. L., Dorland R. B. Response of cultured mammalian cells to the exotoxins of Pseudomonas aeruginosa and Corynebacterium diphtheriae: differential cytotoxicity. Can J Microbiol. 1977 Feb;23(2):183–189. doi: 10.1139/m77-026. [DOI] [PubMed] [Google Scholar]
  15. Sandvig K., Olsnes S. Diphtheria toxin entry into cells is facilitated by low pH. J Cell Biol. 1980 Dec;87(3 Pt 1):828–832. doi: 10.1083/jcb.87.3.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Stephen J. Anthrax toxin. Pharmacol Ther. 1981;12(3):501–513. doi: 10.1016/0163-7258(81)90095-4. [DOI] [PubMed] [Google Scholar]
  17. Vasil M. L., Iglewski B. H. Comparative toxicities of diphtherial toxin and Pseudomonas aeruginosa exotoxin A: evidence for different cell receptors. J Gen Microbiol. 1978 Oct;108(2):333–337. doi: 10.1099/00221287-108-2-333. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES