This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Waterfield, N.
Right arrow Articles by Ffrench-Constant, R. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Waterfield, N.
Right arrow Articles by Ffrench-Constant, R. H.
Agricola
Right arrow Articles by Waterfield, N.
Right arrow Articles by Ffrench-Constant, R. H.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2001, p. 5017-5024, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5017-5024.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Oral Toxicity of Photorhabdus luminescens W14 Toxin Complexes in Escherichia coli

Nicholas Waterfield,1 Andrea Dowling,1 Sadhana Sharma,1 Phillip J. Daborn,1 Ursula Potter,2 and Richard H. Ffrench-Constant1,*

Department of Biology and Biochemistry1 and Centre for Electron Optical Studies,2 University of Bath, Bath BA2 7AY, United Kingdom

Received 21 May 2001/Accepted 9 August 2001

Previous attempts to express the toxin complex genes of Photorhabdus luminescens W14 in Escherichia coli have failed to reconstitute their oral toxicity to the model insect Manduca sexta. Here we show that the combination of three genes, tcdA, tcdB, and tccC, is essential for oral toxicity to M. sexta when expression in E. coli is used. Further, when transcription from native toxin complex gene promoters is used, maximal toxicity in E. coli cultures is associated with the addition of mitomycin C to the growth medium. In contrast, the expression of tcdAB (or the homologous tcaABC operon) with no recombinant tccC homolog in a different P. luminescens strain, K122, is sufficient to confer oral toxicity on this strain, which is otherwise not orally toxic. We therefore infer that P. luminescens K122 carries a functional tccC-like homolog within its own genome, a hypothesis supported by Southern analysis. Recombinant toxins from both P. luminescens K122 and E. coli were purified as high-molecular-weight particulate preparations. Transmission electron micrograph (TEM) images of these particulate preparations showed that the expression of tcdAB (either with or without tccC) in E. coli produces visible ~25-nm-long complexes with a head and tail-like substructure. These data are consistent with a model whereby TcdAB constitutes the majority of the complex visible under TEM and TccC either is a toxin itself or is an activator of the complex. The implications for the potential mode of action of the toxin complex genes are discussed.

* Corresponding author. Mailing address: Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, United Kingdom. Phone: 44 1225 826261. Fax: 44 1225 826779. E-mail: bssrfc{at}bath.ac.uk.

Applied and Environmental Microbiology, November 2001, p. 5017-5024, Vol. 67, No. 11
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.11.5017-5024.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Zhao, R., Han, R., Qiu, X., Yan, X., Cao, L., Liu, X. (2008). Cloning and Heterologous Expression of Insecticidal-Protein-Encoding Genes from Photorhabdus luminescens TT01 in Enterobacter cloacae for Termite Control. Appl. Environ. Microbiol. 74: 7219-7226 [Abstract] [Full Text]  
  • Hares, M. C., Hinchliffe, S. J., Strong, P. C. R., Eleftherianos, I., Dowling, A. J., ffrench-Constant, R. H., Waterfield, N. (2008). The Yersinia pseudotuberculosis and Yersinia pestis toxin complex is active against cultured mammalian cells. Microbiology 154: 3503-3517 [Abstract] [Full Text]  
  • Nunez-Valdez, M. E., Calderon, M. A., Aranda, E., Hernandez, L., Ramirez-Gama, R. M., Lina, L., Rodriguez-Segura, Z., Gutierrez, M. d. C., Villalobos, F. J. (2008). Identification of a Putative Mexican Strain of Serratia entomophila Pathogenic against Root-Damaging Larvae of Scarabaeidae (Coleoptera). Appl. Environ. Microbiol. 74: 802-810 [Abstract] [Full Text]  
  • Sergeant, M., Baxter, L., Jarrett, P., Shaw, E., Ousley, M., Winstanley, C., Morgan, J. A. W. (2006). Identification, Typing, and Insecticidal Activity of Xenorhabdus Isolates from Entomopathogenic Nematodes in United Kingdom Soil and Characterization of the xpt Toxin Loci. Appl. Environ. Microbiol. 72: 5895-5907 [Abstract] [Full Text]  
  • Tan, B., Jackson, T. A., Hurst, M. R. H. (2006). Virulence of Serratia Strains against Costelytra zealandica. Appl. Environ. Microbiol. 72: 6417-6418 [Abstract] [Full Text]  
  • Tennant, S. M., Skinner, N. A., Joe, A., Robins-Browne, R. M. (2005). Homologues of Insecticidal Toxin Complex Genes in Yersinia enterocolitica Biotype 1A and Their Contribution to Virulence. Infect. Immun. 73: 6860-6867 [Abstract] [Full Text]  
  • Derzelle, S., Turlin, E., Duchaud, E., Pages, S., Kunst, F., Givaudan, A., Danchin, A. (2004). The PhoP-PhoQ Two-Component Regulatory System of Photorhabdus luminescens Is Essential for Virulence in Insects. J. Bacteriol. 186: 1270-1279 [Abstract] [Full Text]  
  • Marokhazi, J., Waterfield, N., LeGoff, G., Feil, E., Stabler, R., Hinds, J., Fodor, A., ffrench-Constant, R. H. (2003). Using a DNA Microarray To Investigate the Distribution of Insect Virulence Factors in Strains of Photorhabdus Bacteria. J. Bacteriol. 185: 4648-4656 [Abstract] [Full Text]  
  • Sergeant, M., Jarrett, P., Ousley, M., Morgan, J. A. W. (2003). Interactions of Insecticidal Toxin Gene Products from Xenorhabdus nematophilus PMFI296. Appl. Environ. Microbiol. 69: 3344-3349 [Abstract] [Full Text]  
  • Bowen, D. J., Rocheleau, T. A., Grutzmacher, C. K., Meslet, L., Valens, M., Marble, D., Dowling, A., ffrench-Constant, R., Blight, M. A. (2003). Genetic and biochemical characterization of PrtA, an RTX-like metalloprotease from Photorhabdus. Microbiology 149: 1581-1591 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»