Cysteine residues in the Vif protein of human immunodeficiency virus type 1 are essential for viral infectivity.
The infectivity factor of human immunodeficiency virus type 1 (HIV-1), Vif, contains two cysteine residues which are highly conserved among animal lentiviruses. We introduced substitutions of leucine for cysteine residues in the vif gene of a full-length HIV-1 clone to analyze their roles in viral infection. Mutant viruses containing substitutions in either Cys-114, Cys-133, or both displayed a vif-negative infection phenotype similar to that of an isogeneic vif deletion mutant, namely, a cell-dependent complete to partial loss of infectivity. The vif defect could be complemented by cotransfection of mutant viral DNA with a Vif expression vector, and there was no evidence that recombination contributed to the repair of the vif deficiency. The viral protein profile, as determined by immunoblotting, in cells infected with cysteine substitution mutants and that in wild-type virus were similar, including the presence of the 23-kDa Vif polypeptide. In addition, immunoblotting with an antiserum directed against the carboxyl terminus of gp41 revealed that gp41 was intact in cells infected with either wild-type or vif mutant HIV-1, excluding that Vif cleaves the C terminus of gp41. Our results indicate that the cysteines in HIV-1 Vif are critical for Vif function in viral infectivity.
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Molecular Virology Laboratory, St. Luke's/Roosevelt Hospital Center, New York, New York 10019.
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Chao
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Volsky
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D000595:Amino Acid Sequence; D001483:Base Sequence; D003545:Cysteine / Q000235:genetics; D016346:Gene Products, vif / Q000096:biosynthesis / Q000235:genetics*; D005816:Genetic Complementation Test; D016655:HIV Core Protein p24 / Q000096:biosynthesis; D015700:HIV Envelope Protein gp41 / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D011499:Protein Processing, Post-Translational; D012689:Sequence Homology, Nucleic Acid; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000382:microbiology*; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000096:biosynthesis; D014774:Virulence; D054320:vif Gene Products, Human Immunodeficiency Virus
D016346:Gene Products, vif; D016655:HIV Core Protein p24; D015700:HIV Envelope Protein gp41; D014764:Viral Proteins; D054320:vif Gene Products, Human Immunodeficiency Virus; D003545:Cysteine