Characterization of the fusion domain of the human immunodeficiency virus type 1 envelope glycoprotein gp41.
The human immunodeficiency virus transmembrane glycoprotein gp41 has at its amino terminus a strongly hydrophobic stretch of 28 amino acids flanked by a highly conserved series of polar amino acids. To investigate the role in syncytium formation of the hydrophobic amino terminus of gp41 and the polar border of this hydrophobic region, we introduced eight single-amino acid substitutions and one double-amino acid substitution in the amino-terminal 31 amino acids of gp41. The mutant envelope glycoproteins were expressed from two distinct human immunodeficiency virus type 1 envelope glycoprotein expression vectors; the effects of the mutations on syncytium formation, envelope glycoprotein transport, secretion, and CD4 receptor-binding were analyzed. Results showed that polar substitutions throughout the hydrophobic amino terminus of gp41 greatly reduced or blocked syncytium formation mediated by the human immunodeficiency virus type 1 envelope glycoproteins, as did nonconservative mutations in the polar border of the hydrophobic amino terminus. Mutations at gp41 amino acids 15, 26, and 29 also significantly increased the extent of gp120 secretion into the extracellular medium. None of the mutations detectably affected envelope glycoprotein processing or envelope glycoprotein binding to CD4.
Freed
E O
EO
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Myers
D J
DJ
Risser
R
R
D000595:Amino Acid Sequence; D015704:CD4 Antigens / Q000032:analysis / Q000276:immunology; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D015700:HIV Envelope Protein gp41 / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006367:HeLa Cells / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D014760:Viral Fusion Proteins / Q000235:genetics*
D015704:CD4 Antigens; D015700:HIV Envelope Protein gp41; D014760:Viral Fusion Proteins