V3 loop region of the HIV-1 gp120 envelope protein is essential for virus infectivity.
The mechanism by which HIV-1 mediates cell fusion and penetrates target cells, subsequent to receptor (CD4) binding, is not well understood. However, neutralizing antibodies, which recognize the principal neutralizing determinants of the gp120 envelope protein (the V3 loop region, residues 296 to 331), have been shown to effectively block cell fusion and virus infectivity independent of the initial gp120-CD4 binding. To investigate the role of the V3 loop in an HIV infection, a series of site-specific mutations were introduced into the HIV-1 envelope gene. Specifically, each residue (312 to 315) in the strongly conserved tetrapeptide sequence, GPGR, which is positioned in the center of the V3 loop domain was individually altered. The processing, transport, and CD4 binding properties of the mutant envelope proteins were comparable to those of the wild-type protein, however, none of the mutants were able to form syncytia in the HeLa-T4 assay. Molecular HIV-1 clones containing mutations altering the G312, G314, or R315 residues produced noninfectious virions, whereas a clone with a P313A mutation was found to be infectious. These results demonstrate that certain V3 loop mutations can be lethal and clearly indicate that this region of the HIV-1 gp120 protein is essential for virus infectivity.
Ivanoff
L A
LA
SmithKline Beecham Pharmaceuticals, Department of Antiinfectives, King of Prussia, Pennsylvania 19406-0939.
Dubay
J W
JW
Morris
J F
JF
Roberts
S J
SJ
Gutshall
L
L
Sternberg
E J
EJ
Hunter
E
E
Matthews
T J
TJ
Petteway
S R
SR
D000595:Amino Acid Sequence; D001483:Base Sequence; D015704:CD4 Antigens / Q000378:metabolism; D002459:Cell Fusion; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D015497:HIV-1 / Q000472:pathogenicity*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D013329:Structure-Activity Relationship; D014779:Virus Replication
D015704:CD4 Antigens; D015699:HIV Envelope Protein gp120; D009838:Oligodeoxyribonucleotides