Targeting of Shiga toxin B-subunit to retrograde transport route in association with detergent-resistant membranes.
In HeLa cells, Shiga toxin B-subunit is transported from the plasma membrane to the endoplasmic reticulum, via early endosomes and the Golgi apparatus, circumventing the late endocytic pathway. We describe here that in cells derived from human monocytes, i.e., macrophages and dendritic cells, the B-subunit was internalized in a receptor-dependent manner, but retrograde transport to the biosynthetic/secretory pathway did not occur and part of the internalized protein was degraded in lysosomes. These differences correlated with the observation that the B-subunit associated with Triton X-100-resistant membranes in HeLa cells, but not in monocyte-derived cells, suggesting that retrograde targeting to the biosynthetic/secretory pathway required association with specialized microdomains of biological membranes. In agreement with this hypothesis we found that in HeLa cells, the B-subunit resisted extraction by Triton X-100 until its arrival in the target compartments of the retrograde pathway, i.e., the Golgi apparatus and the endoplasmic reticulum. Furthermore, destabilization of Triton X-100-resistant membranes by cholesterol extraction potently inhibited B-subunit transport from early endosomes to the trans-Golgi network, whereas under the same conditions, recycling of transferrin was not affected. Our data thus provide first evidence for a role of lipid asymmetry in membrane sorting at the interface between early endosomes and the trans-Golgi network.
Falguières
T
T
Unité Mixte de Recherche 144 Institut Curie/ Centre National de la Recherche Scientifique, F-75248 Paris Cedex 05, France.
Mallard
F
F
Baron
C
C
Hanau
D
D
Lingwood
C
C
Goud
B
B
Salamero
J
J
Johannes
L
L
D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002469:Cell Separation; D002784:Cholesterol / Q000378:metabolism; D003713:Dendritic Cells / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism; D005434:Flow Cytometry; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000378:metabolism; D017830:Octoxynol / Q000494:pharmacology; D021122:Protein Subunits; D021381:Protein Transport / Q000502:physiology*; D022621:Shiga Toxin / Q000378:metabolism*; D014281:Trihexosylceramides / Q000378:metabolism
D003902:Detergents; D021122:Protein Subunits; D014281:Trihexosylceramides; C018549:globotriaosylceramide; D022621:Shiga Toxin; D017830:Octoxynol; D002784:Cholesterol