Cytoplasmic domain requirement for incorporation of a foreign envelope protein into vesicular stomatitis virus.
Incorporation of human immunodeficiency virus type 1 (HIV-1) envelope proteins into vesicular stomatitis virus (VSV) particles was studied in a system that allows expressed envelope proteins to rescue phenotypically a temperature-sensitive mutant of VSV (tsO45). This mutant exhibits defective transport of its own envelope glycoprotein (G) and can be rescued by simultaneous expression of wild-type G protein from cDNA. We report here that a hybrid HIV-1-VSV protein containing the extracellular and transmembrane domains of the HIV-1 envelope protein fused to the cytoplasmic domain of VSV G protein was able to rescue the tsO45 mutant lacking the G protein, while the wild-type HIV-1 envelope protein was not. The VSV(HIV) pseudotypes obtained infected only CD4+ cells and were neutralized specifically by anti-HIV-1 sera. Our results indicate that the cytoplasmic tail of the VSV glycoprotein contains an independent signal capable of directing a foreign protein into VSV particles. The VSV(HIV) pseudotypes generated here were prepared in the absence of HIV-1 and should be useful for identifying molecules that block HIV-1 entry.
Owens
R J
RJ
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510-8023.
Rose
J K
JK
D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology; D002459:Cell Fusion; D005455:Fluorescent Antibody Technique; D015686:Gene Products, env / Q000302:isolation & purification / Q000378:metabolism*; D015699:HIV Envelope Protein gp120 / Q000302:isolation & purification / Q000378:metabolism; D015700:HIV Envelope Protein gp41 / Q000302:isolation & purification / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013329:Structure-Activity Relationship; D014721:Vesicular stomatitis Indiana virus / Q000254:growth & development*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism*
C028391:G protein, vesicular stomatitis virus; D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins