Intracellular transport of a variant surface glycoprotein in Trypanosoma brucei.
Trypanosome variant surface glycoproteins (VSGs) have a novel glycan-phosphatidylinositol membrane anchor, which is cleavable by a phosphatidylinositol-specific phospholipase C. A similar structure serves to anchor some membrane proteins in mammalian cells. Using kinetic and ultrastructural approaches, we have addressed the question of whether this structure directs the protein to the cell surface by a different pathway from the classical one described in other cell types for plasma membrane and secreted glycoproteins. By immunogold labeling on thin cryosections we were able to show that, intracellularly, VSG is associated with the rough endoplasmic reticulum, all Golgi cisternae, and tubulovesicular elements and flattened cisternae, which form a network in the area adjacent to the trans side of the Golgi apparatus. Our data suggest that, although the glycan-phosphatidylinositol anchor is added in the endoplasmic reticulum, VSG is nevertheless subsequently transported along the classical intracellular route for glycoproteins, and is delivered to the flagellar pocket, where it is integrated into the surface coat. Treatment of trypanosomes with 1 microM monensin had no effect on VSG transport, although dilation of the trans-Golgi stacks and lysosomes occurred immediately. Incubation of trypanosomes at 20 degrees C, a treatment that arrests intracellular transport from the trans-Golgi region to the cell surface in mammalian cells, caused the accumulation of VSG molecules in structures of the trans-Golgi network, and retarded the incorporation of newly synthesized VSG into the surface coat.
Duszenko
M
M
Rockefeller University, New York, New York 10021.
Ivanov
I E
IE
Ferguson
M A
MA
Plesken
H
H
Cross
G A
GA
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002451:Cell Compartmentation; D006017:Glycolipids / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D008985:Monensin / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D014346:Trypanosoma brucei brucei / Q000378:metabolism* / Q000648:ultrastructure; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000378:metabolism*
D006017:Glycolipids; D014643:Variant Surface Glycoproteins, Trypanosoma; D008985:Monensin