In vitro mutagenesis identifies a region within the envelope gene of the human immunodeficiency virus that is critical for infectivity.
Site-specific mutagenesis was used to introduce amino acid substitutions at the asparagine codons of four conserved potential N-linked glycosylation sites within the gp120 envelope protein of human immunodeficiency virus (HIV). One of these alterations resulted in the production of noninfectious virus particles. The amino acid substitution did not interfere with the synthesis, processing, and stability of the env gene polypeptides gp120 and gp41 or the binding of gp120 to its cellular receptor, the CD4 (T4) molecule. Vaccinia virus recombinants containing wild-type or mutant HIV env genes readily induced syncytia in CD4+ HeLa cells. These results suggest that alterations involving the second conserved domain of the HIV gp120 may interfere with an essential early step in the virus replication cycle other than binding to the CD4 receptor. In long-term cocultures of a T4+ lymphocyte cell line and colon carcinoma cells producing the mutant virus, revertant infectious virions were detected. Molecular characterization of two revertant proviral clones revealed the presence of the original mutation as well as a compensatory amino acid change in another region of HIV gp120.
Willey
R L
RL
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Smith
D H
DH
Lasky
L A
LA
Theodore
T S
TS
Earl
P L
PL
Moss
B
B
Capon
D J
DJ
Martin
M A
MA
D000945:Antigens, Differentiation, T-Lymphocyte / Q000502:physiology; D002459:Cell Fusion; D004252:DNA Mutational Analysis; D005814:Genes, Viral; D006678:HIV / Q000235:genetics / Q000472:pathogenicity*; D009024:Morphogenesis; D011991:Receptors, Virus / Q000502:physiology; D013601:T-Lymphocytes / Q000382:microbiology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000502:physiology; D014771:Virion / Q000502:physiology; D014779:Virus Replication
D000945:Antigens, Differentiation, T-Lymphocyte; D011991:Receptors, Virus; D014759:Viral Envelope Proteins