Role of the conserved dipeptide Gly75 and Cys76 on HIV-1 Vpr function.
Vpr is one of the accessory proteins encoded by the HIV-1 genome. Several interesting features associated with Vpr include incorporation into virus particles, ability to oligomerize, localization in the nucleus, and positive effect on virus production and replication. In order to understand the structure-function relationship of Vpr, we have analyzed the role of the Gly75 and Cys76 (GC) residues which are highly conserved in HIV-1 Vpr and in Vpr and Vpx of HIV-2/SIV. We have generated several substitution mutants involving this dipeptide and have evaluated for expression, stability, nuclear localization, and virion incorporation of Vpr. Our data demonstrate that the GC residues are not essential for virion incorporation and nuclear localization of Vpr. Serine substitution for Cys, however, restricted the localization of Vpr in the cytoplasm without affecting the Gag-directed incorporation of Vpr into virus-like particles. Interestingly, the cysteine-substituted mutants showed altered stability in comparison to the wild type, and substitution mutants for glycine showed minimal effect on stability. These results indicate that the glycine and cysteine do not play a role in nuclear localization or virion incorporation properties of Vpr and further suggest that these two functions of Vpr may not be interdependent.
Mahalingam
S
S
Institute of Biotechnology, Jefferson Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Collman
R G
RG
Patel
M
M
Monken
C E
CE
Srinivasan
A
A
D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000737:chemistry; D017124:Conserved Sequence / Q000235:genetics*; D003545:Cysteine / Q000502:physiology; D003593:Cytoplasm / Q000737:chemistry; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004151:Dipeptides / Q000235:genetics / Q000502:physiology*; D016336:Gene Products, vpr / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D016334:Genes, vpr / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D005998:Glycine / Q000502:physiology; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D009154:Mutation / Q000502:physiology; D014616:Vaccinia virus / Q000235:genetics; D014764:Viral Proteins; D014771:Virion; D014779:Virus Replication / Q000235:genetics*; D054325:vpr Gene Products, Human Immunodeficiency Virus
D004151:Dipeptides; D016336:Gene Products, vpr; D014764:Viral Proteins; D054325:vpr Gene Products, Human Immunodeficiency Virus; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D003545:Cysteine; D005998:Glycine