| Autor: |
Genisset, Christophe, Galeotti, Cesira L., Lupetti, Pietro, Mercati, David, Skibinski, David A. G., Barone, Silvia, Battistutta, Roberto, de Bernard, Marina, Telford, John L. |
| Zdroj: |
Infection and Immunity; March 2006, Vol. 74 Issue: 3 p1786-1794, 9p |
| Abstrakt: |
ABSTRACTMost Helicobacter pyloristrains secrete a toxin (VacA) that causes massive vacuolization of target cells and which is a major virulence factor of H. pylori. The VacA amino-terminal region is required for the induction of vacuolization. The aim of the present study was a deeper understanding of the critical role of the N-terminal regions that are protected from proteolysis when VacA interacts with artificial membranes. Using a counterselection system, we constructed an H. pyloristrain, SPM 326-Δ49-57, that produces a mutant toxin with a deletion of eight amino acids in one of these protected regions. VacA Δ49-57 was correctly secreted by H. pyloribut failed to oligomerize and did not have any detectable vacuolating cytotoxic activity. However, the mutant toxin was internalized normally and stained the perinuclear region of HeLa cells. Moreover, the mutant toxin exhibited a dominant negative effect, completely inhibiting the vacuolating activity of wild-type VacA. This loss of activity was correlated with the disappearance of oligomers in electron microscopy. These findings indicate that the deletion in VacA Δ49-57 disrupts the intermolecular interactions required for the oligomerization of the toxin. |
| Databáze: |
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