Biosynthesis and processing of human immunodeficiency virus type 1 envelope glycoproteins: effects of monensin on glycosylation and transport.
When human immunodeficiency virus type 1 envelope glycoproteins were expressed in 293 cells by using a recombinant adenovirus expression vector, the envelope precursor (gp160) was initially glycosylated by cotranslational addition of N-linked high-mannose oligosaccharide units to the protein backbone and then cleaved to gp120 and gp41. The subunits gp120 and gp41 were then further modified by the addition of fucose, galactose, and sialic acid, resulting in glycoproteins containing a mixture of hybrid and complex oligosaccharide side chains. A fraction of glycosylated gp160 that escaped cleavage was further modified by the terminal addition of fucose and galactose, but the addition of sialic acid did not occur, consistent with the notion that it is compartmentalized separately from the gp120 envelope protein. Processing and transport of gp160 were blocked by the monovalent ionophore monensin, which at high concentrations (25 microM and above) was a potent inhibitor of the endoproteolytic cleavage of gp160; at lower concentrations (1 to 10 microM), it selectively blocked the secondary glycosylation steps so that smaller products were produced. Monensin (1 microM) treatment also resulted in a reduction in syncytium formation, which was observed when recombinant infected cells were cocultivated with CD4-bearing HeLa cells. The infectivity of human immunodeficiency virus type 1 was also reduced by monensin treatment, a decrease that may be due to incompletely glycosylated forms of gp120 that have a lower affinity for the CD4 receptor.
Dewar
R L
RL
Division of Molecular Virology and Immunology, Georgetown University School of Medicine, D.C. 20007.
Vasudevachari
M B
MB
Natarajan
V
V
Salzman
N P
NP
D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D006031:Glycosylation; D015488:HIV Antigens / Q000096:biosynthesis / Q000378:metabolism; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D015497:HIV-1 / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008985:Monensin / Q000494:pharmacology*; D011233:Precipitin Tests; D012191:Retroviridae Proteins / Q000096:biosynthesis* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000378:metabolism
D015488:HIV Antigens; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins; D008985:Monensin