Genetic Rearrangements in the Pathogenicity Locus of Clostridium Difficile Strain 8864--implications for Transcription, Expression and Enzymatic Activity of Toxins A and B

Mol Gen Genet. 1998 May;258(3):222-32. doi: 10.1007/s004380050726.


The pathogenicity locus (PaLoc) of Clostridium difficile isolate 8864 was investigated to locate genetic rearrangements that would explain the exceptional pathogenicity of this particular isolate. Two major changes were defined: an insertion of 1.1 kb between the two genes tcdA and tcdE, coding for the enterotoxin and an accessory protein of unknown function, respectively, and a deletion of 5.9 kb encompassing the 3' ends of tcdA and tcdC. Transcription of the tcdA-E genes is severely affected by both rearrangements, explaining the demonstrated complete lack of TcdA polypeptide. We present a model of coordinate, growth-related transcription of the tcdA-E genes that confirms our previous findings in strain 10463. Recombinant TcdA-8864 had UDP-glucose-glucosyltransferase activity, proving that the N-terminal 698 amino acids of the polypeptide represent the catalytic domain. However, this truncated TcdA molecule lacks a ligand and translocation domain. To assess the catalytic domain of TcdB-8864, the sequence of the 5' end of its gene was determined. TcdB-8864 shows high homology to TcdB-1470 but lower homology to TcdB-10463 within this domain. This fits well with the altered glucosylation specificity of TcdB-8864 (Rac1, Rap2 and Ra1). Having defined the variations of transcription, expression and enzymatic activity of toxins A and B, implications for the pathogenic potential of strain 8864 are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Toxins / chemistry*
  • Clostridium difficile / genetics
  • Clostridium difficile / pathogenicity*
  • GTP-Binding Proteins / metabolism
  • Gene Expression Regulation, Bacterial / genetics*
  • Genes, Bacterial / genetics
  • Glucosyltransferases / metabolism
  • Glycoside Hydrolases / metabolism
  • Glycosylation
  • Molecular Sequence Data
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Transcription, Genetic / genetics


  • Bacterial Proteins
  • Bacterial Toxins
  • Recombinant Proteins
  • Glucosyltransferases
  • Glycoside Hydrolases
  • GTP-Binding Proteins