Truncation of the human immunodeficiency virus type 1 transmembrane glycoprotein cytoplasmic domain blocks virus infectivity.
Human immunodeficiency virus type 1 contains a transmembrane glycoprotein with an unusually long cytoplasmic domain. To determine the role of this domain in virus replication, a series of single nucleotide changes that result in the insertion of premature termination codons throughout the cytoplasmic domain has been constructed. These mutations delete from 6 to 192 amino acids from the carboxy terminus of gp41 and do not affect the amino acid sequence of the regulatory proteins encoded by rev and tat. The effects of these mutations on glycoprotein biosynthesis and function as well as on virus infectivity have been examined in the context of a glycoprotein expression vector and the viral genome. All of the mutant glycoproteins were synthesized, processed, and transported to the cell surface in a manner similar to that of the wild-type glycoprotein. With the exception of mutants that remove the membrane anchor domain, all of the mutant glycoproteins retained the ability to cause fusion of CD4-bearing cells. However, deletion of more than 19 amino acids from the C terminus of gp41 blocked the ability of mutant virions to infect cells. This defect in virus infectivity appeared to be due at least in part to a failure of the virus to efficiently incorporate the truncated glycoprotein. Similar data were obtained for mutations in two different env genes and two different target cell lines. These results indicate that the cytoplasmic domain of gp41 plays a critical role during virus assembly and entry in the life cycle of human immunodeficiency virus type 1.
Dubay
J W
JW
Department of Microbiology, University of Alabama, Birmingham 35294.
Roberts
S J
SJ
Hahn
B H
BH
Hunter
E
E
D000818:Animals; D001692:Biological Transport; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology; D002459:Cell Fusion; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D003062:Codon; D004274:DNA, Recombinant; D015686:Gene Products, env / Q000235:genetics*; D015752:Genes, env / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics / Q000472:pathogenicity*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D014176:Protein Biosynthesis; D011487:Protein Conformation; D013728:Terminator Regions, Genetic / Q000235:genetics; D014162:Transfection; D014760:Viral Fusion Proteins; D014774:Virulence
D003062:Codon; D004274:DNA, Recombinant; D015686:Gene Products, env; D014760:Viral Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase