Effects of brefeldin A on the processing of viral envelope glycoproteins in murine erythroleukemia cells.
This paper documents the effects of brefeldin A (BFA) on the processing and transport of viral envelope glycoproteins in a retrovirus-transformed murine erythroleukemia (MEL) cell line. BFA is a fungal metabolite that disrupts intracellular membrane traffic at the endoplasmic reticulum (ER)-Golgi complex junction. In MEL cells, BFA inhibited the processing of the newly synthesized precursor, gPr90env, of the murine leukemia virus envelope protein, gp70, and curtailed the budding of virions into the culture medium by blocking the transport of this protein out of the ER. The block resulted in the intracellular accumulation of gPr90env and two putative products of its processing (78 and 66 kDa). The results of endoglycosidase (endo) H and D digestion of the viral glycoproteins in the presence and absence of BFA indicated that (i) there was no glycoprotein processing during the first approximately 2 h of the BFA block; (ii) active Golgi enzymes relocated to the ER in approximately 2 h during BFA treatment, resulting in the production of partially endo H-resistant forms of the spleen focus-forming virus glycoprotein, gp55 (in controls, this glycoprotein was generally retained in the ER as an endo H-sensitive entity); and (iii) proteolytic processing of gPr90env to gp70 occurred prior to the acquisition of endo H resistance and at approximately the same time as endo D sensitivity (i.e. in a cis Golgi compartment). In control cells, the spleen focus-forming virus glycoprotein, gp55, underwent turnover with a half-life of approximately 5 h. In contrast, its turnover was considerably slower during BFA treatment (t 1/2 = approximately 20 h), suggesting that transport of gp55 out of the ER was required for its degradation or that BFA afforded it protection from proteolysis within the ER.
Ulmer
J B
JB
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510.
Palade
G E
GE
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D020126:Brefeldin A; D002451:Cell Compartmentation; D003517:Cyclopentanes / Q000494:pharmacology*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015686:Gene Products, env / Q000378:metabolism*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D066298:In Vitro Techniques; D009052:Leukemia Virus, Murine / Q000378:metabolism*; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D013155:Spleen Focus-Forming Viruses / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000378:metabolism*
D003517:Cyclopentanes; D015686:Gene Products, env; D014759:Viral Envelope Proteins; D020126:Brefeldin A