Structural and functional characterization of the human immunodeficiency virus rev protein.
Expression of the human immunodeficiency virus (HIV) rev protein is required for expression of virus structural proteins. Site-directed mutagenesis was used to localize regions important for Rev function. We found that proteins with single amino acid substitution mutations concentrated within the amino termini and midportion of Rev were for the most part nonfunctional. Indirect immunofluorescence revealed that Rev was localized predominantly in the nucleolus. However, a deletion mutant that lacked the basic stretch of amino acids comprising residues Arg-Arg-Arg-Arg-Trp accumulated in the cytoplasm and was no longer functional. Consistent with this observation, a beta-galactosidase fusion protein containing this basic rich peptide at its amino termini was targeted to the nucleus. These observations indicate that the HIV rev protein has a distinct nuclear localization sequence and suggest that Rev-mediated regulation of gene expression involves nuclear events.
Perkins
A
A
Department of Molecular Oncology, Roche Institute of Molecular Biology, Roche Research Center, Nutley, New Jersey 07110.
Cochrane
A W
AW
Ruben
S M
SM
Rosen
C A
CA
D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000032:analysis; D002478:Cells, Cultured; D005455:Fluorescent Antibody Technique; D015694:Gene Products, rev; D005814:Genes, Viral; D006678:HIV / Q000032:analysis* / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D013347:Subcellular Fractions / Q000032:analysis; D014764:Viral Proteins / Q000302:isolation & purification*; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D014764:Viral Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus