Mutational analysis of the cleavage sequence of the human immunodeficiency virus type 1 envelope glycoprotein precursor gp160.
The envelope glycoproteins of the human immunodeficiency virus (HIV) type 1 are synthesized as a precursor molecule, gp160, which is cleaved to generate the two mature envelope glycoproteins, gp120 and gp41. The cleavage reaction, which is mediated by a host protease, occurs at a sequence highly conserved in retroviral envelope glycoprotein precursors. We have investigated the sequence requirements for this cleavage reaction by introducing four single-amino-acid changes into the glutamic acid-lysine-arginine sequence immediately amino terminal to the site of cleavage. We have also examined the effects of these mutations on the syncytium formation induced by HIV envelope glycoproteins. Our results indicate that a glutamic acid to glycine change at gp120 amino acid 516, a lysine to isoleucine change at amino acid 517, and an arginine to lysine change at amino acid 518 affect neither gp160 cleavage nor syncytium formation. The results obtained with the arginine to lysine change at amino acid 518 differ significantly from the results obtained with the same mutation at the envelope precursor cleavage site of a murine leukemia virus (E. O. Freed, and R. Risser, J. Virol. 61:2852-2856, 1987). An arginine to threonine mutation at gp120 amino acid 518, the terminal residue of gp120, abolishes both gp160 cleavage and syncytium formation. These findings demonstrate that despite its highly conserved nature, the basic pair of amino acids at the site of gp160 cleavage is not absolutely required for proper envelope glycoprotein processing. This report also supports the idea that cleavage of gp160 is required for activation of the HIV envelope fusion function.
Freed
E O
EO
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Myers
D J
DJ
Risser
R
R
D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D015686:Gene Products, env / Q000032:analysis / Q000235:genetics*; D005814:Genes, Viral; D005822:Genetic Vectors; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells / Q000166:cytology; D006801:Humans; D009154:Mutation; D014162:Transfection; D015678:Viral Structural Proteins / Q000235:genetics*
D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D015678:Viral Structural Proteins