Document 0106
 DOCN  M9470106
 TI    Biochemical mechanism of HIV-I Vpr function. Specific interaction with a
       cellular protein.
 DT    9409
 AU    Zhao LJ; Mukherjee S; Narayan O; Department of Microbiology, Molecular
       Genetics, and Immunology,; University of Kansas Medical Center, Kansas
       City 66160-7424.
 SO    J Biol Chem. 1994 Jun 3;269(22):15577-82. Unique Identifier : AIDSLINE
       MED/94253140
 AB    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.
 DE    Amino Acid Sequence  Animal  Base Sequence  Binding Sites  Carrier
       Proteins/ISOLATION & PURIF/*METABOLISM  Cell Line  Cell Nucleus
       Cloning, Molecular  Comparative Study  DNA Primers  Escherichia
       coli/*METABOLISM  Gene Products, vpr/BIOSYNTHESIS/ISOLATION &
       PURIF/*METABOLISM  *Genes, vpr  Genome, Viral  Hela Cells  Human
       HIV-1/GENETICS/*METABOLISM  Molecular Sequence Data  Mutagenesis,
       Site-Directed  Oligonucleotides, Antisense  Polymerase Chain Reaction
       Recombinant Proteins/BIOSYNTHESIS/ISOLATION & PURIF/METABOLISM
       Repetitive Sequences, Nucleic Acid  Restriction Mapping  Sequence
       Homology, Amino Acid  Support, U.S. Gov't, P.H.S.  Transfection  JOURNAL
       ARTICLE

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