The p21 Rho-activating toxin cytotoxic necrotizing factor 1 is endocytosed by a clathrin-independent mechanism and enters the cytosol by an acidic-dependent membrane translocation step.
Cytotoxic necrotizing factor 1 (CNF1), a protein produced by pathogenic strains of Escherichia coli, activates the p21 Rho-GTP-binding protein, inducing a profound reorganization of the actin cytoskeleton. CNF1 binds to its cell surface receptor on HEp-2 cells with high affinity (K(d) = 20 pM). In HEp-2 cells the action of CNF1 is not blocked in the presence of filipin, a drug described to reduce cholera toxin internalization by the caveolae-like mechanism. Moreover, HEp-2 cells, which express a dominant negative form of proteins that impair the formation of clathrin coated-vesicles and internalization of transferrin (Eps15, dynamin or intersectin-Src homology 3), are still sensitive to CNF1. In this respect, the endocytosis of CNF1 is similar to the plant toxin ricin. However, unlike ricin toxin, CNF1 does not cross the Golgi apparatus and requires an acidic cell compartment to transfer its enzymatic activity into the cytosol in a manner similar to that required by diphtheria toxin. As shown for diphtheria toxin, the pH-dependent membrane translocation step of CNF1 could be mimicked at the level of the plasma membrane by a brief exposure to a pH of </=5.2. CNF1 is the first bacterial toxin described that uses both a clathrin-independent endocytic mechanism and an acidic-dependent membrane translocation step in its delivery of the catalytic domain to the cell cytosol.
Contamin
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Institut National de la Santé et de la Recherche Médicale, Unité 452, Faculté de Médecine, 06107 Nice Cedex 2, France.
Galmiche
A
A
Doye
A
A
Flatau
G
G
Benmerah
A
A
Boquet
P
P
D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism* / Q000493:pharmacokinetics; D001665:Binding Sites; D001692:Biological Transport; D020134:Catalytic Domain; D051242:Caveolin 1; D022461:Caveolins; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D018699:Coated Vesicles / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D003603:Cytotoxins / Q000235:genetics / Q000378:metabolism* / Q000493:pharmacokinetics; D004285:Dogs; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D029968:Escherichia coli Proteins; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D014407:Tumor Cells, Cultured
D001427:Bacterial Toxins; D051242:Caveolin 1; D022461:Caveolins; D002966:Clathrin; D003603:Cytotoxins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; C060296:cytotoxic necrotizing factor type 1