Analysis of the Neurotoxin Complex Genes in Clostridium Botulinum A1-A4 and B1 Strains: BoNT/A3, /Ba4 and /B1 Clusters Are Located Within Plasmids

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2007 Dec 7

PMID 18060065

Citations 81

Authors

Theresa J Smith

Karen K Hill

Brian T Foley

John C Detter

A Christine Munk

David C Bruce

Norman A Doggett

Leonard A Smith

James D Marks

Gary Xie

Thomas S Brettin

Affiliations

Soon will be listed here.

Abstract

Background: Clostridium botulinum and related clostridial species express extremely potent neurotoxins known as botulinum neurotoxins (BoNTs) that cause long-lasting, potentially fatal intoxications in humans and other mammals. The amino acid variation within the BoNT is used to categorize the species into seven immunologically distinct BoNT serotypes (A-G) which are further divided into subtypes. The BoNTs are located within two generally conserved gene arrangements known as botulinum progenitor complexes which encode toxin-associated proteins involved in toxin stability and expression.

Methodology/principal Findings: Because serotype A and B strains are responsible for the vast majority of human botulism cases worldwide, the location, arrangement and sequences of genes from eight different toxin complexes representing four different BoNT/A subtypes (BoNT/A1-Ba4) and one BoNT/B1 strain were examined. The bivalent Ba4 strain contained both the BoNT/A4 and BoNT/bvB toxin clusters. The arrangements of the BoNT/A3 and BoNT/A4 subtypes differed from the BoNT/A1 strains and were similar to those of BoNT/A2. However, unlike the BoNT/A2 subtype, the toxin complex genes of BoNT/A3 and BoNT/A4 were found within large plasmids and not within the chromosome. In the Ba4 strain, both BoNT toxin clusters (A4 and bivalent B) were located within the same 270 kb plasmid, separated by 97 kb. Complete genomic sequencing of the BoNT/B1 strain also revealed that its toxin complex genes were located within a 149 kb plasmid and the BoNT/A3 complex is within a 267 kb plasmid.

Conclusions/significance: Despite their size differences and the BoNT genes they contain, the three plasmids containing these toxin cluster genes share significant sequence identity. The presence of partial insertion sequence (IS) elements, evidence of recombination/gene duplication events, and the discovery of the BoNT/A3, BoNT/Ba4 and BoNT/B1 toxin complex genes within plasmids illustrate the different mechanisms by which these genes move among diverse genetic backgrounds of C. botulinum.

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References

Nielsen H, KROGH A . Prediction of signal peptides and signal anchors by a hidden Markov model. Proc Int Conf Intell Syst Mol Biol. 1998; 6:122-30. View

Dineen S, Bradshaw M, Johnson E . Neurotoxin gene clusters in Clostridium botulinum type A strains: sequence comparison and evolutionary implications. Curr Microbiol. 2003; 46(5):345-52. DOI: 10.1007/s00284-002-3851-1. View

Zhou Y, Sugiyama H, Nakano H, Johnson E . The genes for the Clostridium botulinum type G toxin complex are on a plasmid. Infect Immun. 1995; 63(5):2087-91. PMC: 173270. DOI: 10.1128/iai.63.5.2087-2091.1995. View

Sakaguchi Y, Hayashi T, Kurokawa K, Nakayama K, Oshima K, Fujinaga Y . The genome sequence of Clostridium botulinum type C neurotoxin-converting phage and the molecular mechanisms of unstable lysogeny. Proc Natl Acad Sci U S A. 2005; 102(48):17472-7. PMC: 1283531. DOI: 10.1073/pnas.0505503102. View

Bateman A, Birney E, Cerruti L, Durbin R, Etwiller L, Eddy S . The Pfam protein families database. Nucleic Acids Res. 2001; 30(1):276-80. PMC: 99071. DOI: 10.1093/nar/30.1.276. View

Webb R, Smith T, Wright P, Montgomery V, Meagher M, Smith L . Protection with recombinant Clostridium botulinum C1 and D binding domain subunit (Hc) vaccines against C and D neurotoxins. Vaccine. 2007; 25(21):4273-82. DOI: 10.1016/j.vaccine.2007.02.081. View

Mulder N, Apweiler R, Attwood T, Bairoch A, Barrell D, Bateman A . The InterPro Database, 2003 brings increased coverage and new features. Nucleic Acids Res. 2003; 31(1):315-8. PMC: 165493. DOI: 10.1093/nar/gkg046. View

Boeckmann B, Bairoch A, Apweiler R, Blatter M, Estreicher A, Gasteiger E . The SWISS-PROT protein knowledgebase and its supplement TrEMBL in 2003. Nucleic Acids Res. 2003; 31(1):365-70. PMC: 165542. DOI: 10.1093/nar/gkg095. View

Carver T, Rutherford K, Berriman M, Rajandream M, Barrell B, Parkhill J . ACT: the Artemis Comparison Tool. Bioinformatics. 2005; 21(16):3422-3. DOI: 10.1093/bioinformatics/bti553. View

10.

Bruggemann H, Gottschalk G . Insights in metabolism and toxin production from the complete genome sequence of Clostridium tetani. Anaerobe. 2006; 10(2):53-68. DOI: 10.1016/j.anaerobe.2003.08.001. View

11.

Edgar R . MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 2004; 32(5):1792-7. PMC: 390337. DOI: 10.1093/nar/gkh340. View

12.

Lindstrom M, Korkeala H . Laboratory diagnostics of botulism. Clin Microbiol Rev. 2006; 19(2):298-314. PMC: 1471988. DOI: 10.1128/CMR.19.2.298-314.2006. View

13.

Gill D . Bacterial toxins: a table of lethal amounts. Microbiol Rev. 1982; 46(1):86-94. PMC: 373212. DOI: 10.1128/mr.46.1.86-94.1982. View

14.

Harris M, Clark J, Ireland A, Lomax J, Ashburner M, Foulger R . The Gene Ontology (GO) database and informatics resource. Nucleic Acids Res. 2003; 32(Database issue):D258-61. PMC: 308770. DOI: 10.1093/nar/gkh036. View

15.

Haft D, Loftus B, Richardson D, Yang F, Eisen J, Paulsen I . TIGRFAMs: a protein family resource for the functional identification of proteins. Nucleic Acids Res. 2000; 29(1):41-3. PMC: 29844. DOI: 10.1093/nar/29.1.41. View

16.

Lamanna C, GLASSMAN H . The Isolation of Type B Botulinum Toxin. J Bacteriol. 1947; 54(5):575-84. PMC: 526594. DOI: 10.1128/jb.54.5.575-584.1947. View

17.

Hill K, Smith T, Helma C, Ticknor L, Foley B, Svensson R . Genetic diversity among Botulinum Neurotoxin-producing clostridial strains. J Bacteriol. 2006; 189(3):818-32. PMC: 1797315. DOI: 10.1128/JB.01180-06. View

18.

Kurtz S, Phillippy A, Delcher A, Smoot M, Shumway M, Antonescu C . Versatile and open software for comparing large genomes. Genome Biol. 2004; 5(2):R12. PMC: 395750. DOI: 10.1186/gb-2004-5-2-r12. View

19.

Raffestin S, Marvaud J, Cerrato R, Dupuy B, Popoff M . Organization and regulation of the neurotoxin genes in Clostridium botulinum and Clostridium tetani. Anaerobe. 2006; 10(2):93-100. DOI: 10.1016/j.anaerobe.2004.01.001. View

20.

Lowe T, Eddy S . tRNAscan-SE: a program for improved detection of transfer RNA genes in genomic sequence. Nucleic Acids Res. 1997; 25(5):955-64. PMC: 146525. DOI: 10.1093/nar/25.5.955. View