Ganglioside GD3 Traffics from the trans-Golgi Network to Plasma Membrane by a Rab11-independent and Brefeldin A-insensitive Exocytic Pathway

• Published in: The Journal of biological chemistry 2004-11, Vol.279 (46), p.47610-47618
• Main Authors: Pilar Maria Crespo, Ramiro Iglesias-BartolomÃ, Jose Luis Daniotti
• Format: Article
• Language: English
• Subjects: Animals; Biological Transport - physiology; Brefeldin A - metabolism; Cell Membrane - metabolism; Ceramides - metabolism; CHO Cells; Cricetinae; Detergents - metabolism; Exocytosis - physiology; Gangliosides - metabolism; Glycosylphosphatidylinositols - metabolism; Membrane Glycoproteins - metabolism; Octoxynol - metabolism; Propanolamines - metabolism; Protein Synthesis Inhibitors - metabolism; Pyrrolidines - metabolism; rab GTP-Binding Proteins - genetics; rab GTP-Binding Proteins - metabolism; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Sphingomyelins - metabolism; trans-Golgi Network - metabolism; Vesicular stomatitis virus; Viral Envelope Proteins - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M407181200

Gangliosides, complex glycosphingolipids containing sialic acids, have been found to reside in glycosphingolipid-enriched microdomains (GEM) at the plasma membrane. They are synthesized in the lumen of the Golgi complex and appear unable to translocate from the lumenal toward the cytosolic surface of Golgi membrane to access the monomeric lipid transport. As a consequence, they can only leave the Golgi complex via the lumenal surface of transport vesicles. In this work we analyzed the exocytic transport of the disialo ganglioside GD3 from trans -Golgi network (TGN) to plasma membrane in CHO-K1 cells by immunodetection of endogenously synthesized GD3. We found that ganglioside GD3, unlike another luminal membrane-bounded lipid (glycosylphosphatidylinositol-anchored protein), did not partition into GEM domains in the Golgi complex and trafficked from TGN to plasma membrane by a brefeldin A-insensitive exocytic pathway. Moreover, a dominant negative form of Rab11, which prevents exit of vesicular stomatitis virus glycoprotein from the Golgi complex, did not influence the capacity of GD3 to reach the cell surface. Our results strongly support the notion that most ganglioside GD3 traffics from the TGN to the plasma membrane by a non-conventional vesicular pathway where lateral membrane segregation of vesicular stomatitis virus glycoprotein (non-GEM resident) and glycosylphosphatidylinositol-anchored proteins (GEM resident) from GD3 is required before exiting TGN.