Mutations in the cytoplasmic domain of human immunodeficiency virus type 1 transmembrane protein impair the incorporation of Env proteins into mature virions.
In-frame stop codons were introduced into the coding region of human immunodeficiency virus type 1 (HIV-1) transmembrane protein (gp41). Truncation of 147 amino acids from the carboxyl terminus of gp41 (TM709) significantly decreased the stability and cell surface expression of the viral Env proteins, while truncation of 104 amino acids (TM752) did not. Truncation of 43 or more amino acids from the carboxyl terminus of gp41 generated mutant viruses which were noninfectious in several human CD4+ T lymphoid cell lines and fresh peripheral blood mononuclear cells. Analysis of the noninfectious mutant virions revealed significantly reduced incorporation of the Env proteins compared with the wild-type virions. Comparable amounts of Env proteins were detected on the surfaces of wild-type- and TM752-transfected cells, suggesting that the structures of gp41 required for efficient incorporation of Env proteins were disrupted in mutant TM752. Truncation of the last 12 amino acids (TM844) from the carboxyl terminus of gp41 did not significantly affect the assembly and release of virions or the incorporation of Env proteins into mature virions. However, the TM844 virus had dramatically decreased infectivity compared with the wild-type virus. This suggests that the cytoplasmic domain of gp41 also plays a role in other steps of virus replication.
Yu
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Department of Cancer Biology, Harvard School of Public Health, Boston, Massachusetts 02115.
Yuan
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McLane
M F
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Lee
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Essex
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D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D001692:Biological Transport; D002460:Cell Line; D003062:Codon; D004252:DNA Mutational Analysis; D005455:Fluorescent Antibody Technique; D015686:Gene Products, env / Q000378:metabolism*; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D015497:HIV-1 / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011499:Protein Processing, Post-Translational; D013728:Terminator Regions, Genetic / Q000235:genetics; D014162:Transfection; D014771:Virion / Q000254:growth & development / Q000378:metabolism*; D014774:Virulence
D000911:Antibodies, Monoclonal; D003062:Codon; D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D008565:Membrane Proteins