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M9470106.TXT
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1994-07-02
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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
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).