Glycosylphosphatidylinositol-anchored Proteins and Actin Cytoskeleton Modulate Chloride Transport by Channels Formed by the Helicobacter pylori Vacuolating Cytotoxin VacA in HeLa Cells

Gauthier, Nils C.; Ricci, Vittorio; Gounon, Pierre; Doye, Anne; Tauc, Michel; Poujeol, Philippe; Boquet, Patrice

doi:10.1074/jbc.m312040200

Cited by 42 publications

(57 citation statements)

References 42 publications

(86 reference statements)

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“…5G). A similar phenomenon has been reported to occur in the case of the Helicobacter pylori vaculating cytotoxin VacA (53). In cytochalasin D-treated cells manifesting these membrane blebs, the cellular uptake of VP22-GFP was significantly reduced (Fig.…”

Section: Lipid Raft Dependence Of Cellular Uptake Of Vp22-gfp-dy-supporting

confidence: 51%

“…Upon treatment of HeLa cells with cytochalasin D, an actin-depolymerizing reagent, actin filament disruption occurred, with consequent large membrane bleb (Fig. 5G) or protrusion (53) formation on the cell surface. Virtually all cell-surface VP22-GFP signals appeared to be co-localized with membrane blebs induced by cytochalasin D (Fig.…”

Section: Lipid Raft Dependence Of Cellular Uptake Of Vp22-gfp-dy-mentioning

confidence: 99%

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Cellular Internalization of Green Fluorescent Protein Fused with Herpes Simplex Virus Protein VP22 via a Lipid Raft-mediated Endocytic Pathway Independent of Caveolae and Rho Family GTPases but Dependent on Dynamin and Arf6

Nishi

Saigo

2007

Journal of Biological Chemistry

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VP22 is a structural protein of the herpes simplex virus and has been reported to possess unusual trafficking properties. Here we examined the mechanism of cellular uptake of VP22 using a fusion protein between the C-terminal half of VP22 and green fluorescent protein (GFP). Adsorption of VP22-GFP onto a cell surface required heparan sulfate proteoglycans and basic amino acids, in particular, Arg-164 of VP22. Inhibitor treatment, RNA interference, expression of dominant-negative mutant genes, and confocal microscopy all indicated that VP22-GFP enters cells through an endocytic pathway independent of clathrin and caveolae but dependent on dynamin and Arf6 activity. As with CD59 (a lipid raft marker), cell-surface VP22-GFP signals were resistant to Triton X-100 treatment but only partially overlapped cell-surface CD59 signals. Furthermore, unlike other lipid raft-mediated endocytic pathways, no Rho family GTPase was required for VP22-GFP internalization. Internalized VP22 initially entered early endosomes and then moved to lysosomes and possibly recycling endosomes.

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Section: Lipid Raft Dependence Of Cellular Uptake Of Vp22-gfp-dy-supporting

confidence: 51%

Section: Lipid Raft Dependence Of Cellular Uptake Of Vp22-gfp-dy-mentioning

confidence: 99%

Cellular Internalization of Green Fluorescent Protein Fused with Herpes Simplex Virus Protein VP22 via a Lipid Raft-mediated Endocytic Pathway Independent of Caveolae and Rho Family GTPases but Dependent on Dynamin and Arf6

Nishi

Saigo

2007

Journal of Biological Chemistry

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“…Lipid rafts that contain cellular cholesterol have been shown to be utilized by VacA for binding and entry, hence exhibiting intoxication activity (23,29,40,44,46). Wunder et al recently reported that H. pylori absorbs cellular cholesterol and transforms it into glucosylated derivatives to evade phagocytosis during infection (63).…”

Section: Discussionmentioning

confidence: 99%

“…Several types of evidence suggest that VacA intoxication relies on cellular lipid rafts, which are dynamic microdomains containing cholesterol, sphingolipids, and glycosylphosphatidylinositol-anchored proteins in the exoplasmic leaflet of the lipid bilayer and are implicated in cellular signaling, membrane transport and trafficking, and membrane curvature (for reviews, see references 26 and 51). First, cellular cholesterol is required for VacA cytotoxicity in cultured gastric cells (23,40,46), and VacA migrates with tubulovesicular structures that contain glycosylphosphatidylinositol-anchored proteins to enter cells for its full vacuolating activity (23,29,44). Furthermore, using a cold detergent extraction method, VacA has been isolated primarily on detergent-resistant membranes (DRMs) that are generally considered to be fractions of rafts (23,40,46).…”

mentioning

confidence: 99%

“…First, cellular cholesterol is required for VacA cytotoxicity in cultured gastric cells (23,40,46), and VacA migrates with tubulovesicular structures that contain glycosylphosphatidylinositol-anchored proteins to enter cells for its full vacuolating activity (23,29,44). Furthermore, using a cold detergent extraction method, VacA has been isolated primarily on detergent-resistant membranes (DRMs) that are generally considered to be fractions of rafts (23,40,46). It has been noted that cholesterol-rich microdomains are also utilized by other bacterial toxins, including cholera toxin (38) and aerolysin (1), for entry or oligomerization.…”

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confidence: 99%

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Cholesterol Depletion ReducesHelicobacter pyloriCagA Translocation and CagA-Induced Responses in AGS Cells

et al. 2008

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Infection with Helicobacter pylori cagA-positive strains is associated with gastritis, ulcerations, and gastric cancer. CagA is translocated into infected epithelial cells by a type IV secretion system and can be tyrosine phosphorylated, inducing signal transduction and motogenic responses in epithelial cells. Cellular cholesterol, a vital component of the membrane, contributes to membrane dynamics and functions and is important in VacA intoxication and phagocyte evasion during H. pylori infection. In this investigation, we showed that cholesterol extraction by methyl-␤-cyclodextrin reduced the level of CagA translocation and phosphorylation. Confocal microscope visualization revealed that a significant portion of translocated CagA was colocalized with the raft marker GM1 and c-Src during infection. Moreover, GM1 was rapidly recruited into sites of bacterial attachment by live-cell imaging analysis. CagA and VacA were cofractionated with detergent-resistant membranes (DRMs), suggesting that the distribution of CagA and VacA is associated with rafts in infected cells. Upon cholesterol depletion, the distribution shifted to non-DRMs. Accordingly, the CagA-induced hummingbird phenotype and interleukin-8 induction were blocked by cholesterol depletion. Raft-disrupting agents did not influence bacterial adherence but did significantly reduce internalization activity in AGS cells. Together, these results suggest that delivery of CagA into epithelial cells by the bacterial type IV secretion system is mediated in a cholesterol-dependent manner.

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Toxins in the Endosomes

Reig

Goot

2006

Endosomes

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Glycosylphosphatidylinositol-anchored Proteins and Actin Cytoskeleton Modulate Chloride Transport by Channels Formed by the Helicobacter pylori Vacuolating Cytotoxin VacA in HeLa Cells

Cited by 42 publications

References 42 publications

Cellular Internalization of Green Fluorescent Protein Fused with Herpes Simplex Virus Protein VP22 via a Lipid Raft-mediated Endocytic Pathway Independent of Caveolae and Rho Family GTPases but Dependent on Dynamin and Arf6

Cellular Internalization of Green Fluorescent Protein Fused with Herpes Simplex Virus Protein VP22 via a Lipid Raft-mediated Endocytic Pathway Independent of Caveolae and Rho Family GTPases but Dependent on Dynamin and Arf6

Cholesterol Depletion ReducesHelicobacter pyloriCagA Translocation and CagA-Induced Responses in AGS Cells

Toxins in the Endosomes

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