Studies on the role of the V3 loop in human immunodeficiency virus type 1 envelope glycoprotein function.
Mutations within the principal neutralizing determinant (the V3 loop) of the HIV-1 surface envelope glycoprotein gp120 block or greatly reduce the ability of the HIV-1 envelope glycoprotein to induce cell fusion in CD4+ HeLa T4 cells while keeping its CD4 binding ability. However, when either cysteine or both cysteines forming the V3 disulfide bridge were mutated, the resultant glycoprotein could not mediate cell fusion, undergo proteolytic processing, or bind CD4. To investigate the role that the V3 loop plays in gp160 processing and CD4 binding, we deleted the entire V3 loop region of the HIV-1 env gene. The resultant glycoprotein could not mediate cell fusion in the HeLa T4 cell line and no proteolytic processing of gp160 or CD4 binding could be detected. To test whether any domain of the V3 loop is involved in attaining the proper envelope glycoprotein conformation required for proteolytic processing and CD4 binding, we introduced a series of deletions into the coding region of the V3 loop. Most of the residues within the V3 loop could be removed while retaining gp160 processing and CD4 binding. Our results indicate that the cysteines that form the V3 loop or the disulfide bond itself are important for proper envelope glycoprotein folding and processing. Because many of the mutants constructed in this study do not contain the type-specific neutralizing determinant of HIV-1, they may be potential reagents to bind group-specific neutralizing antibodies or to elicit a group-specific neutralizing response against HIV-1.
Chiou
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Department of Biochemistry, Cancer Research University of Wisconsin-Madison 53706.
Freed
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Panganiban
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Kenealy
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D000595:Amino Acid Sequence; D001483:Base Sequence; D015704:CD4 Antigens / Q000378:metabolism; D003001:Cloning, Molecular; D004279:DNA, Viral; D015726:Giant Cells / Q000382:microbiology; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000502:physiology*; D015497:HIV-1 / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008561:Membrane Fusion / Q000502:physiology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000502:physiology*; D011233:Precipitin Tests; D011485:Protein Binding; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D017384:Sequence Deletion
D015704:CD4 Antigens; D004279:DNA, Viral; D015699:HIV Envelope Protein gp120; C059762:HIV envelope protein gp120 (305-321); D010446:Peptide Fragments