The carboxy-terminal domain is essential for stability and not for virion incorporation of HIV-1 Vpr into virus particles.
Vpr is one of the auxiliary gene products encoded by HIV-1 genome. Vpr is a 14-kDa protein and exhibits several interesting characteristics including incorporation into virus particles, oligomerization, localization in the nucleus, and positive regulation of virus replication in primary cells. In an effort to define the structure-function relationship of Vpr, the role of the C-terminus of Vpr was investigated. Site-specific mutagenesis involving deletion, insertion, and substitution of residues at the C-terminus was utilized to generate variants of Vpr. Mutations introduced at the C-terminus affected properties of Vpr in different ways: (i) Vpr containing amino acids 1-72 showed the virion incorporation phenotype, indicating that the C-terminus is not essential for this function, (ii) the C-terminus contributes to the stability of Vpr, and (iii) substitution mutagenesis involving the basic residues showed stability similar to that of wild type, indicating the lack of involvement of these residues in this biochemical property of Vpr. The data generated in this study and our early mutagenic analyses on Vpr suggest that domains noncontiguous in primary sequence contribute to the stability of Vpr through overall conformation of the protein.
Mahalingam
S
S
Department of Microbiology and Immunology, Jefferson Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Patel
M
M
Collman
R G
RG
Srinivasan
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002264:Carboxylic Acids; D017931:DNA Primers; D016336:Gene Products, vpr / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D013329:Structure-Activity Relationship; D014771:Virion / Q000378:metabolism; D054325:vpr Gene Products, Human Immunodeficiency Virus
D002264:Carboxylic Acids; D017931:DNA Primers; D016336:Gene Products, vpr; D054325:vpr Gene Products, Human Immunodeficiency Virus