Effect of lysosomotropic amines on the secretory pathway and on the recycling of the asialoglycoprotein receptor in human hepatoma cells.
We studied the intracellular transport of secretory and membrane proteins in the human hepatoma cell line HepG-2 infected with vesicular stomatitis virus. Cells were pulse-labeled in the presence of [35S]methionine and chased in the presence of the lysosomotropic agent primaquine. At a concentration of 0.3 mM primaquine effectively inhibited the secretion of albumin and, to a lesser extent, that of orosomucoid and transferrin. The drug also prevented the budding of virus particles at the cell surface. The intracellular transport to the Golgi complex of the membrane protein VSV-G was not affected by primaquine as it acquires resistance to endo-beta-N-acetylglucosaminidase H at the same rate as in control cells. Addition of primaquine at various times after the initiation of the chase period indicates that the effect of primaquine occurs just before secretion. In confirmation of the biochemical data, immunocytochemical localization of albumin in cells treated with NH4Cl demonstrated that albumin accumulated in vesicles at the trans side of the Golgi complex. The effect of primaquine on secretion was also compared with its effect on receptor recycling. The dose-response characteristics of the effect of primaquine on receptor recycling are identical to those of the effects on protein secretion and virus budding. These results indicate that both processes involve the same transport mechanism, and/or that they occur via at least one identical intracellular compartment.
Strous
G J
GJ
Du Maine
A
A
Zijderhand-Bleekemolen
J E
JE
Slot
J W
JW
Schwartz
A L
AL
D037263:Asialoglycoprotein Receptor; D001693:Biological Transport, Active / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000648:ultrastructure; D002460:Cell Line; D002472:Cell Transformation, Viral / Q000187:drug effects; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms; D008175:Lung Neoplasms / Q000378:metabolism / Q000648:ultrastructure; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D008565:Membrane Proteins / Q000096:biosynthesis; D009363:Neoplasm Proteins / Q000378:metabolism; D011319:Primaquine / Q000494:pharmacology*; D011971:Receptors, Immunologic / Q000187:drug effects / Q000378:metabolism*; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism
D037263:Asialoglycoprotein Receptor; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D011971:Receptors, Immunologic; D014764:Viral Proteins; D011319:Primaquine