This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Perelle, S Articles by Popoff, M R Search for Related Content PubMed PubMed Citation Articles by Perelle, S Articles by Popoff, M R

Characterization of Clostridium perfringens iota-toxin genes and expression in Escherichia coli.

Laboratoire des Toxines Microbiennes, Institut Pasteur, Paris, France.

ABSTRACT

The iota toxin which is produced by Clostridium perfringens type E, is a binary toxin consisting of two independent polypeptides: Ia, which is an ADP-ribosyltransferase, and Ib, which is involved in the binding and internalization of the toxin into the cell. Two degenerate oligonucleotide probes deduced from partial amino acid sequence of each component of C. spiroforme toxin, which is closely related to the iota toxin, were used to clone three overlapping DNA fragments containing the iota-toxin genes from C. perfringens type E plasmid DNA. Two genes, in the same orientation, coding for Ia (387 amino acids) and Ib (875 amino acids) and separated by 243 noncoding nucleotides were identified. A predicted signal peptide was found for each component, and the secreted Ib displays two domains, the propeptide (172 amino acids) and the mature protein (664 amino acids). The Ia gene has been expressed in Escherichia coli and C. perfringens, under the control of its own promoter. The recombinant polypeptide obtained was recognized by Ia antibodies and ADP-ribosylated actin. The expression of the Ib gene was obtained in E. coli harboring a recombinant plasmid encompassing the putative promoter upstream of the Ia gene and the Ia and Ib genes. Two residues which have been found to be involved in the NAD+ binding site of diphtheria and pseudomonas toxins are conserved in the predicted Ia sequence (Glu-14 and Trp-19). The predicted amino acid Ib sequence shows 33.9% identity with and 54.4% similarity to the protective antigen of the anthrax toxin complex. In particular, the central region of Ib, which contains a predicted transmembrane segment (Leu-292 to Ser-308), presents 45% identity with the corresponding protective antigen sequence which is involved in the translocation of the toxin across the cell membrane.

This article has been cited by other articles:

• Fehr, D., Burr, S. E., Gibert, M., d'Alayer, J., Frey, J., Popoff, M. R. (2007). Aeromonas Exoenzyme T of Aeromonas salmonicida Is a Bifunctional Protein That Targets the Host Cytoskeleton. J. Biol. Chem. 282: 28843-28852 [Abstract] [Full Text]  
• Keel, M. K., Songer, J. G. (2006). The Comparative Pathology of Clostridium difficile-associated Disease. Vet Pathol 43: 225-240 [Abstract] [Full Text]  
• Pituch, H., Kreft, D., Obuch-Woszczatynski, P., Wultanska, D., Meisel-Mikolajczyk, F., Luczak, M., van Belkum, A. (2005). Clonal Spread of a Clostridium difficile Strain with a Complete Set of Toxin A, Toxin B, and Binary Toxin Genes among Polish Patients with Clostridium difficile-Associated Diarrhea. J. Clin. Microbiol. 43: 472-475 [Abstract] [Full Text]  
• Barth, H., Aktories, K., Popoff, M. R., Stiles, B. G. (2004). Binary Bacterial Toxins: Biochemistry, Biology, and Applications of Common Clostridium and Bacillus Proteins. Microbiol. Mol. Biol. Rev. 68: 373-402 [Abstract] [Full Text]  
• Nagahama, M., Yamaguchi, A., Hagiyama, T., Ohkubo, N., Kobayashi, K., Sakurai, J. (2004). Binding and Internalization of Clostridium perfringens Iota-Toxin in Lipid Rafts. Infect. Immun. 72: 3267-3275 [Abstract] [Full Text]  
• Haug, G., Aktories, K., Barth, H. (2004). The Host Cell Chaperone Hsp90 Is Necessary for Cytotoxic Action of the Binary Iota-Like Toxins. Infect. Immun. 72: 3066-3068 [Abstract] [Full Text]  
• Goncalves, C., Decre, D., Barbut, F., Burghoffer, B., Petit, J.-C. (2004). Prevalence and Characterization of a Binary Toxin (Actin-Specific ADP-Ribosyltransferase) from Clostridium difficile. J. Clin. Microbiol. 42: 1933-1939 [Abstract] [Full Text]  
• Hale, M. L., Marvaud, J.-C., Popoff, M. R., Stiles, B. G. (2004). Detergent-Resistant Membrane Microdomains Facilitate Ib Oligomer Formation and Biological Activity of Clostridium perfringens Iota-Toxin. Infect. Immun. 72: 2186-2193 [Abstract] [Full Text]  
• Haug, G., Leemhuis, J., Tiemann, D., Meyer, D. K., Aktories, K., Barth, H. (2003). The Host Cell Chaperone Hsp90 Is Essential for Translocation of the Binary Clostridium botulinum C2 Toxin into the Cytosol. J. Biol. Chem. 278: 32266-32274 [Abstract] [Full Text]  
• Vidal, C., Geny, B., Melle, J., Jandrot-Perrus, M., Fontenay-Roupie, M. (2002). Cdc42/Rac1-dependent activation of the p21-activated kinase (PAK) regulates human platelet lamellipodia spreading: implication of the cortical-actin binding protein cortactin. Blood 100: 4462-4469 [Abstract] [Full Text]  
• Marvaud, J.-C., Stiles, B. G., Chenal, A., Gillet, D., Gibert, M., Smith, L. A., Popoff, M. R. (2002). Clostridium perfringens Iota Toxin. MAPPING OF THE Ia DOMAIN INVOLVED IN DOCKING WITH Ib AND CELLULAR INTERNALIZATION. J. Biol. Chem. 277: 43659-43666 [Abstract] [Full Text]  
• Nagahama, M., Nagayasu, K., Kobayashi, K., Sakurai, J. (2002). Binding Component of Clostridium perfringens Iota-Toxin Induces Endocytosis in Vero Cells. Infect. Immun. 70: 1909-1914 [Abstract] [Full Text]  
• Knapp, O., Benz, R., Gibert, M., Marvaud, J. C., Popoff, M. R. (2002). Interaction of Clostridium perfringens Iota-Toxin with Lipid Bilayer Membranes. DEMONSTRATION OF CHANNEL FORMATION BY THE ACTIVATED BINDING COMPONENT Ib AND CHANNEL BLOCK BY THE ENZYME COMPONENT Ia. J. Biol. Chem. 277: 6143-6152 [Abstract] [Full Text]  
• Pannucci, J., Okinaka, R. T., Sabin, R., Kuske, C. R. (2002). Bacillus anthracis pXO1 Plasmid Sequence Conservation among Closely Related Bacterial Species. J. Bacteriol. 184: 134-141 [Abstract] [Full Text]  
• Blocker, D., Behlke, J., Aktories, K., Barth, H. (2001). Cellular Uptake of the Clostridium perfringens Binary Iota-Toxin. Infect. Immun. 69: 2980-2987 [Abstract] [Full Text]  
• Marvaud, J.-C., Smith, T., Hale, M. L., Popoff, M. R., Smith, L. A., Stiles, B. G. (2001). Clostridium perfringens Iota-Toxin: Mapping of Receptor Binding and Ia Docking Domains on Ib. Infect. Immun. 69: 2435-2441 [Abstract] [Full Text]  
• Zdanovsky, A. G., Zdanovskaia, M. V. (2000). Simple and Efficient Method for Heterologous Expression of Clostridial Proteins. Appl. Environ. Microbiol. 66: 3166-3173 [Abstract] [Full Text]  
• Alonso, A., Sanchez, P., Martínez, J. L. (2000). Stenotrophomonas maltophilia D457R Contains a Cluster of Genes from Gram-Positive Bacteria Involved in Antibiotic and Heavy Metal Resistance. Antimicrob. Agents Chemother. 44: 1778-1782 [Abstract] [Full Text]  
• Gibert, M., Petit, L., Raffestin, S., Okabe, A., Popoff, M. R. (2000). Clostridium perfringens Iota-Toxin Requires Activation of Both Binding and Enzymatic Components for Cytopathic Activity. Infect. Immun. 68: 3848-3853 [Abstract] [Full Text]  
• Stiles, B. G., Hale, M. L., Marvaud, J.-C., Popoff, M. R. (2000). Clostridium perfringens Iota Toxin: Binding Studies and Characterization of Cell Surface Receptor by Fluorescence-Activated Cytometry. Infect. Immun. 68: 3475-3484 [Abstract] [Full Text]  
• Nagahama, M., Sakaguchi, Y., Kobayashi, K., Ochi, S., Sakurai, J. (2000). Characterization of the Enzymatic Component of Clostridium perfringens Iota-Toxin. J. Bacteriol. 182: 2096-2103 [Abstract] [Full Text]  
• Eckhardt, M., Barth, H., Blocker, D., Aktories, K. (2000). Binding of Clostridium botulinum C2 Toxin to Asparagine-linked Complex and Hybrid Carbohydrates. J. Biol. Chem. 275: 2328-2334 [Abstract] [Full Text]  
• El Hadj, N. B., Popoff, M. R., Marvaud, J.-C., Payrastre, B., Boquet, P., Geny, B. (1999). G-protein-stimulated Phospholipase D Activity Is Inhibited by Lethal Toxin from Clostridium sordellii in HL-60 Cells. J. Biol. Chem. 274: 14021-14031 [Abstract] [Full Text]  
• Price, L. B., Hugh-Jones, M., Jackson, P. J., Keim, P. (1999). Genetic Diversity in the Protective Antigen Gene of Bacillus anthracis. J. Bacteriol. 181: 2358-2362 [Abstract] [Full Text]  
• Varughese, M., Teixeira, A. V., Liu, S., Leppla, S. H. (1999). Identification of a Receptor-Binding Region within Domain 4 of the Protective Antigen Component of Anthrax Toxin. Infect. Immun. 67: 1860-1865 [Abstract] [Full Text]  
• Billington, S. J., Wieckowski, E. U., Sarker, M. R., Bueschel, D., Songer, J. G., McClane, B. A. (1998). Clostridium perfringens Type E Animal Enteritis Isolates with Highly Conserved, Silent Enterotoxin Gene Sequences. Infect. Immun. 66: 4531-4536 [Abstract] [Full Text]  
• Gasman, S., Chasserot-Golaz, S., Hubert, P., Aunis, D., Bader, M.-F. (1998). Identification of a Potential Effector Pathway for the Trimeric Go Protein Associated with Secretory Granules. Go STIMULATES A GRANULE-BOUND PHOSPHATIDYLINOSITOL 4-KINASE BY ACTIVATING RhoA IN CHROMAFFIN CELLS. J. Biol. Chem. 273: 16913-16920 [Abstract] [Full Text]  
• Barth, H., Hofmann, F., Olenik, C., Just, I., Aktories, K. (1998). The N-Terminal Part of the Enzyme Component (C2I) of the Binary Clostridium botulinum C2 Toxin Interacts with the Binding Component C2II and Functions as a Carrier System for a Rho ADP-Ribosylating C3-Like Fusion Toxin. Infect. Immun. 66: 1364-1369 [Abstract] [Full Text]  
• Gasman, S., Chasserot-Golaz, S., Popoff, M. R., Aunis, D., Bader, M.-F. (1997). Trimeric G Proteins Control Exocytosis in Chromaffin Cells. Go REGULATES THE PERIPHERAL ACTIN NETWORK AND CATECHOLAMINE SECRETION BY A MECHANISM INVOLVING THE SMALL GTP-BINDING PROTEIN Rho. J. Biol. Chem. 272: 20564-20571 [Abstract] [Full Text]  
• Sellman, B. R., Nassi, S., Collier, R. J. (2001). Point Mutations in Anthrax Protective Antigen That Block Translocation. J. Biol. Chem. 276: 8371-8376 [Abstract] [Full Text]  
• Singh, Y., Khanna, H., Chopra, A. P., Mehra, V. (2001). A Dominant Negative Mutant of Bacillus anthracis Protective Antigen Inhibits Anthrax Toxin Action in Vivo. J. Biol. Chem. 276: 22090-22094 [Abstract] [Full Text]