Identification of residues in the N-terminal acidic domain of HIV-1 Vpr essential for virion incorporation.
Vpr is one of the auxiliary proteins encoded by the HIV-1 genome and is selectively incorporated into the virus particle. It has been shown that Vpr incorporation in the virus particle requires only the core protein Gag. In an effort to identify the domains of Vpr which are essential for incorporation into the HIV-1 virion, site-specific mutagenesis of vpr was carried out. Mutation of the highly conserved acidic residues in the N-terminal domain (amino acid positions 17-34) eliminated virion incorporation. These mutations disrupt a predicted amphipathic alpha-helical structure that is highly conserved among Vpr sequences. In contrast, alterations of the conserved cysteine (Cys76), basic domain (Arg87 and Lys95), and other residues (Gln65) did not impair the incorporation of Vpr into virus-like particles directed by HIV-1 Gag. The results presented here suggest that protein-protein interactions mediated through the putative helical domain of Vpr may participate in its incorporation into the virus particle.
Mahalingam
S
S
Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.
Khan
S A
SA
Jabbar
M A
MA
Monken
C E
CE
Collman
R G
RG
Srinivasan
A
A
D000596:Amino Acids; D001483:Base Sequence; D017124:Conserved Sequence; D015870:Gene Expression; D015683:Gene Products, gag / Q000235:genetics; D016336:Gene Products, vpr / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D016334:Genes, vpr / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D015497:HIV-1 / Q000254:growth & development*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology; D017433:Protein Structure, Secondary; D014616:Vaccinia virus / Q000235:genetics; D014771:Virion / Q000502:physiology*; D054325:vpr Gene Products, Human Immunodeficiency Virus
D000596:Amino Acids; D015683:Gene Products, gag; D016336:Gene Products, vpr; D054325:vpr Gene Products, Human Immunodeficiency Virus