High cell sensitivity to Helicobacter pylori VacA toxin depends on a GPI-anchored protein and is not blocked by inhibition of the clathrin-mediated pathway of endocytosis

V. Ricci, A. Galmiche, A. Doye, V. Necchi, E. Solcia, P. Boquet

Research output: Contribution to journalArticle

Abstract

Helicobacter pylori vacuolating toxin (VacA) causes vacuolation in a variety of cultured cell lines, sensitivity to VacA differing greatly, however, among the different cell types. We found that the high sensitivity of HEp-2 cells to VacA was impaired by treating the cells with phosphatidylinositol-specific phospholipase C (PI-PLC) which removes glycosylphosphatidylinositol (GPI)-anchored proteins from the cell surface. Incubation of cells with a cholesterol-sequestering agent, that impairs both structure and function of sphingolipid-cholesterol-rich membrane microdomains ('lipid rafts'), also impaired VacA-induced cell vacuolation. Overexpression into HEp-2 cells of proteins inhibiting clathrin-dependent endocytosis (i.e., a dominant-negative mutant of Eps15, the five tandem Src-homology-3 domains of intersectin, and the K44A dominant-negative mutant of dynamin II) did not affect vacuolation induced by VacA. Nevertheless, F-actin depolymerization, known to block the different types of endocytic mechanisms, strongly impaired VacA vacuolating activity. Taken together, our data suggest that the high cell sensitivity to VacA depends on the presence of one or several GPI-anchored protein(s), intact membrane lipid rafts, and an uptake mechanism via a clathrin-independent endocytic pathway.

Original languageEnglish
Pages (from-to)3897-3909
Number of pages13
JournalMolecular Biology of the Cell
Volume11
Issue number11
Publication statusPublished - 2000

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Glycosylphosphatidylinositols
Clathrin
Endocytosis
Helicobacter pylori
Proteins
Membrane Lipids
Sequestering Agents
Dynamin II
Cholesterol
Membrane Microdomains
Phosphoinositide Phospholipase C
Sphingolipids
src Homology Domains
Actins
Cultured Cells
Membrane Proteins
Cell Line

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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High cell sensitivity to Helicobacter pylori VacA toxin depends on a GPI-anchored protein and is not blocked by inhibition of the clathrin-mediated pathway of endocytosis. / Ricci, V.; Galmiche, A.; Doye, A.; Necchi, V.; Solcia, E.; Boquet, P.

In: Molecular Biology of the Cell, Vol. 11, No. 11, 2000, p. 3897-3909.

Research output: Contribution to journalArticle

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AU - Galmiche, A.

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AU - Solcia, E.

AU - Boquet, P.

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