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. 1995 Jul;61(7):2601–2605. doi: 10.1128/aem.61.7.2601-2605.1995

Binding of the CryIVD Toxin of Bacillus thuringiensis subsp. israelensis to Larval Dipteran Midgut Proteins

F Feldmann, A Dullemans, C Waalwijk
PMCID: PMC1388492  PMID: 16535074

Abstract

Ligand-blotting experiments on dipteran brush border membrane vesicles (BBMVs) showed binding of CryIVD toxin of Bacillus thuringiensis subsp. israelensis to proteins of 148 kDa in Anopheles stephensi and of 78 kDa in Tipula oleracea, both species being susceptible to CryIVD. Binding of CryIVD with BBMVs of A. stephensi resulted in a stronger signal than with BBMVs of T. oleracea. Likewise, larvae of A. stephensi are 10,000-fold more susceptible to the CryIVD toxin than are larvae of T. oleracea. Binding was also found with six proteins ranging in size from 48 to 110 kDa in BBMVs from the lepidopteran species Manduca sexta, but CryIVD was not toxic for M. sexta larvae. No binding of trypsinated CryIVD to BBMV proteins was observed. With the lepidopteran-specific toxin CryIA(b), no binding to dipteran BBMVs was found. Binding of CryIA(b) to nine different BBMV proteins ranging in size from 71 to 240 kDa was observed in M. sexta. The major binding signal was observed with a protein of 240 kDa for CryIA(b).

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Selected References

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  1. Aronson A. I., Han E. S., McGaughey W., Johnson D. The solubility of inclusion proteins from Bacillus thuringiensis is dependent upon protoxin composition and is a factor in toxicity to insects. Appl Environ Microbiol. 1991 Apr;57(4):981–986. doi: 10.1128/aem.57.4.981-986.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chilcott C. N., Ellar D. J. Comparative toxicity of Bacillus thuringiensis var. israelensis crystal proteins in vivo and in vitro. J Gen Microbiol. 1988 Sep;134(9):2551–2558. doi: 10.1099/00221287-134-9-2551. [DOI] [PubMed] [Google Scholar]
  3. Dadd R. H. Alkalinity within the midgut of mosquito larvae with alkaline-active digestive enzymes. J Insect Physiol. 1975 Nov;21(11):1847–1853. doi: 10.1016/0022-1910(75)90252-8. [DOI] [PubMed] [Google Scholar]
  4. Dai S. M., Gill S. S. In vitro and in vivo proteolysis of the Bacillus thuringiensis subsp. israelensis CryIVD protein by Culex quinquefasciatus larval midgut proteases. Insect Biochem Mol Biol. 1993 Mar;23(2):273–283. doi: 10.1016/0965-1748(93)90008-g. [DOI] [PubMed] [Google Scholar]
  5. Donovan W. P., Dankocsik C., Gilbert M. P. Molecular characterization of a gene encoding a 72-kilodalton mosquito-toxic crystal protein from Bacillus thuringiensis subsp. israelensis. J Bacteriol. 1988 Oct;170(10):4732–4738. doi: 10.1128/jb.170.10.4732-4738.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Feldmann A. M., Ponnudurai T. Selection of Anopheles stephensi for refractoriness and susceptibility to Plasmodium falciparum. Med Vet Entomol. 1989 Jan;3(1):41–52. doi: 10.1111/j.1365-2915.1989.tb00473.x. [DOI] [PubMed] [Google Scholar]
  7. Ferré J., Real M. D., Van Rie J., Jansens S., Peferoen M. Resistance to the Bacillus thuringiensis bioinsecticide in a field population of Plutella xylostella is due to a change in a midgut membrane receptor. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5119–5123. doi: 10.1073/pnas.88.12.5119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Garczynski S. F., Crim J. W., Adang M. J. Identification of putative insect brush border membrane-binding molecules specific to Bacillus thuringiensis delta-endotoxin by protein blot analysis. Appl Environ Microbiol. 1991 Oct;57(10):2816–2820. doi: 10.1128/aem.57.10.2816-2820.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gould F., Martinez-Ramirez A., Anderson A., Ferre J., Silva F. J., Moar W. J. Broad-spectrum resistance to Bacillus thuringiensis toxins in Heliothis virescens. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7986–7990. doi: 10.1073/pnas.89.17.7986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haider M. Z., Ellar D. J. Analysis of the molecular basis of insecticidal specificity of Bacillus thuringiensis crystal delta-endotoxin. Biochem J. 1987 Nov 15;248(1):197–201. doi: 10.1042/bj2480197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hofmann C., Vanderbruggen H., Höfte H., Van Rie J., Jansens S., Van Mellaert H. Specificity of Bacillus thuringiensis delta-endotoxins is correlated with the presence of high-affinity binding sites in the brush border membrane of target insect midguts. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7844–7848. doi: 10.1073/pnas.85.21.7844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Höfte H., Whiteley H. R. Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev. 1989 Jun;53(2):242–255. doi: 10.1128/mr.53.2.242-255.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ibarra J. E., Federici B. A. Isolation of a relatively nontoxic 65-kilodalton protein inclusion from the parasporal body of Bacillus thuringiensis subsp. israelensis. J Bacteriol. 1986 Feb;165(2):527–533. doi: 10.1128/jb.165.2.527-533.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Martínez-Ramírez A. C., González-Nebauer S., Escriche B., Real M. D. Ligand blot identification of a Manduca sexta midgut binding protein specific to three Bacillus thuringiensis CryIA-type ICPs. Biochem Biophys Res Commun. 1994 Jun 15;201(2):782–787. doi: 10.1006/bbrc.1994.1769. [DOI] [PubMed] [Google Scholar]
  16. McGaughey W. H. Insect Resistance to the Biological Insecticide Bacillus thuringiensis. Science. 1985 Jul 12;229(4709):193–195. doi: 10.1126/science.229.4709.193. [DOI] [PubMed] [Google Scholar]
  17. Oddou P., Hartmann H., Radecke F., Geiser M. Immunologically unrelated Heliothis sp. and Spodoptera sp. midgut membrane-proteins bind Bacillus thuringiensis CryIA(b) delta-endotoxin. Eur J Biochem. 1993 Feb 15;212(1):145–150. doi: 10.1111/j.1432-1033.1993.tb17644.x. [DOI] [PubMed] [Google Scholar]
  18. Ponnudurai T., Lensen A. H., Van Gemert G. J., Bensink M. P., Bolmer M., Meuwissen J. H. Infectivity of cultured Plasmodium falciparum gametocytes to mosquitoes. Parasitology. 1989 Apr;98(Pt 2):165–173. doi: 10.1017/s0031182000062065. [DOI] [PubMed] [Google Scholar]
  19. Rapoport G., Klier A., Billault A., Fargette F., Dedonder R. Construction of a colony bank of E. coli containing hybrid plasmids representative of the Bacillus subtilis 168 genome. Expression of functions harbored by the recombinant plasmids in B. subtilis. Mol Gen Genet. 1979 Oct 3;176(2):239–245. doi: 10.1007/BF00273218. [DOI] [PubMed] [Google Scholar]
  20. Ravoahangimalala O., Charles J. F., Schoeller-Raccaud J. Immunological localization of Bacillus thuringiensis serovar israelensis toxins in midgut cells of intoxicated Anopheles gambiae larvae (Diptera: Culicidae). Res Microbiol. 1993 May;144(4):271–278. doi: 10.1016/0923-2508(93)90011-p. [DOI] [PubMed] [Google Scholar]
  21. Sangadala S., Walters F. S., English L. H., Adang M. J. A mixture of Manduca sexta aminopeptidase and phosphatase enhances Bacillus thuringiensis insecticidal CryIA(c) toxin binding and 86Rb(+)-K+ efflux in vitro. J Biol Chem. 1994 Apr 1;269(13):10088–10092. [PubMed] [Google Scholar]
  22. Thomas W. E., Ellar D. J. Bacillus thuringiensis var israelensis crystal delta-endotoxin: effects on insect and mammalian cells in vitro and in vivo. J Cell Sci. 1983 Mar;60:181–197. doi: 10.1242/jcs.60.1.181. [DOI] [PubMed] [Google Scholar]
  23. Thorne C. B. Transduction in Bacillus thuringiensis. Appl Environ Microbiol. 1978 Jun;35(6):1109–1115. doi: 10.1128/aem.35.6.1109-1115.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vadlamudi R. K., Ji T. H., Bulla L. A., Jr A specific binding protein from Manduca sexta for the insecticidal toxin of Bacillus thuringiensis subsp. berliner. J Biol Chem. 1993 Jun 15;268(17):12334–12340. [PubMed] [Google Scholar]
  25. Van Rie J., Jansens S., Höfte H., Degheele D., Van Mellaert H. Receptors on the brush border membrane of the insect midgut as determinants of the specificity of Bacillus thuringiensis delta-endotoxins. Appl Environ Microbiol. 1990 May;56(5):1378–1385. doi: 10.1128/aem.56.5.1378-1385.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Van Rie J., Jansens S., Höfte H., Degheele D., Van Mellaert H. Specificity of Bacillus thuringiensis delta-endotoxins. Importance of specific receptors on the brush border membrane of the mid-gut of target insects. Eur J Biochem. 1989 Dec 8;186(1-2):239–247. doi: 10.1111/j.1432-1033.1989.tb15201.x. [DOI] [PubMed] [Google Scholar]
  27. Van Rie J., McGaughey W. H., Johnson D. E., Barnett B. D., Van Mellaert H. Mechanism of insect resistance to the microbial insecticide Bacillus thuringiensis. Science. 1990 Jan 5;247(4938):72–74. doi: 10.1126/science.2294593. [DOI] [PubMed] [Google Scholar]
  28. Waalwijk C., Dullemans A. M., van Workum M. E., Visser B. Molecular cloning and the nucleotide sequence of the Mr 28 000 crystal protein gene of Bacillus thuringiensis subsp. israelensis. Nucleic Acids Res. 1985 Nov 25;13(22):8207–8217. doi: 10.1093/nar/13.22.8207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wolfersberger M. G. The toxicity of two Bacillus thuringiensis delta-endotoxins to gypsy moth larvae is inversely related to the affinity of binding sites on midgut brush border membranes for the toxins. Experientia. 1990 May 15;46(5):475–477. doi: 10.1007/BF01954236. [DOI] [PubMed] [Google Scholar]
  30. Yang Y. J., Davies D. M. Trypsin and chymotrypsin during metamorphosis in Aedes aegypti and properties of the chymotrypsin. J Insect Physiol. 1971 Jan;17(1):117–131. doi: 10.1016/0022-1910(71)90048-5. [DOI] [PubMed] [Google Scholar]

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