6533b851fe1ef96bd12a8e33
RESEARCH PRODUCT
A nonsense mutation abrogates production of a functional enterotoxin A in Clostridium difficile toxinotype VIII strains of serogroups F and X.
Christoph Von Eichel-streiberMiklavz GrabnarInes Zec-pirnatInes Zec-pirnatMaja Rupniksubject
Nonsense mutationBlotting WesternMutation MissenseEnterotoxinBiologymedicine.disease_causeMicrobiologyMicrobiologylaw.inventionEnterotoxinsBacterial ProteinslawCatalytic DomainGeneticsmedicineMissense mutationHumansMolecular BiologyGenechemistry.chemical_classificationMutationClostridioides difficileMolecular biologyStop codonPeptide FragmentsRecombinant ProteinsAmino acidchemistryGenes BacterialRecombinant DNAGene Deletiondescription
Clostridium difficile strains of toxinotype VIII from serogroups F and X are described as toxin B-positive, toxin A-negative (TcdB+ A-), although they harbour almost the entire tcdA gene. To identify the reason for the lack of TcdA detection, we analyzed catalytic and ligand domains of TcdA-1470 of the type strain of serogroup F, strain 1470. Using recombinant fragments, the C-terminal immunodominant ligand domain TcdA3-1470, spanning amino acid residues 1694-2711 (corresponding to VPI 10463 sequence), was detected in Western blots. Similar experiments using the recombinant N-terminal catalytic fragment TcdAc1-2-1470 (amino acid positions 1-544) failed. In addition, this fragment showed no glucosylation activity. We determined the size and the position of alterations in the ligand domain tcdA3-1470 by DNA sequencing. Within the N-terminal fragment tcdAc1-2-1470, a nonsense mutation was identified introducing a stop codon at amino acid position 47. Identical mutations were found in the two serogroup X strains 17663 and 10355. The mutation might explain the lack of TcdA production observed in strains of serotypes F and X.
year | journal | country | edition | language |
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1999-09-14 | FEMS microbiology letters |