Biochemical mechanism of HIV-I Vpr function. Specific interaction with a cellular protein.
vpr is an accessory gene of human immunodeficiency virus I (HIV-I). Although unnecessary for viral replication in T cell lines, growing evidence suggests that it is essential for virus replication in monocytes/macrophages and for replication in vivo. We expressed HIV-I vpr in Escherichia coli and purified Vpr by affinity chromatography. ... In a coprecipitation assay, the purified Vpr interacted specifically with a cellular protein designated as Vpr-interacting protein, or RIP. Mutational analysis suggested that this interaction required a domain rich in leucine/isoleucine residues and highly conserved between HIV-I and SIVmac Vprs. During transient expression in mammalian cells, HIV-I Vpr was localized in the nucleus. However, mutational analysis failed to identify in Vpr a typical nuclear localization signal rich in basic amino acid residues. Instead, Vpr nuclear localization seemed to correlate with Vpr interaction with RIP. Mutations in the C-terminal 20-amino acid region containing a cryptic nuclear localization signal did not abolish Vpr nuclear localization or interaction with RIP, whereas point mutations in the leucine/isoleucine-rich domain abolished Vpr interaction with RIP and rendered Vpr unstable during transient expression. These results suggest that RIP may be involved in Vpr function.
Mesh Terms:
Amino Acid Sequence, Animals, Base Sequence, Binding Sites, Carrier Proteins, Cell Line, Cell Nucleus, Cloning, Molecular, DNA Primers, Escherichia coli, Gene Products, vpr, Genes, vpr, Genome, Viral, HIV-1, HeLa Cells, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Oligonucleotides, Antisense, Polymerase Chain Reaction, Recombinant Proteins, Repetitive Sequences, Nucleic Acid, Restriction Mapping, Sequence Homology, Amino Acid, Transfection, vpr Gene Products, Human Immunodeficiency Virus
Amino Acid Sequence, Animals, Base Sequence, Binding Sites, Carrier Proteins, Cell Line, Cell Nucleus, Cloning, Molecular, DNA Primers, Escherichia coli, Gene Products, vpr, Genes, vpr, Genome, Viral, HIV-1, HeLa Cells, Humans, Molecular Sequence Data, Mutagenesis, Site-Directed, Oligonucleotides, Antisense, Polymerase Chain Reaction, Recombinant Proteins, Repetitive Sequences, Nucleic Acid, Restriction Mapping, Sequence Homology, Amino Acid, Transfection, vpr Gene Products, Human Immunodeficiency Virus
J. Biol. Chem.
Date: Jun. 03, 1994
PubMed ID: 8195203
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