Trans-activating rev protein of the human immunodeficiency virus 1 interacts directly and specifically with its target RNA.
The 20-kDa phosphorylated rev protein from human immunodeficiency virus 1 has been shown to transactivate posttranscriptionally the expression of viral structural proteins by selective stabilization and nuclear export of unspliced and incompletely spliced viral mRNA. We could demonstrate in gel-mobility and immunoprecipitation assays that the recombinant rev protein purified from a baculovirus expression system forms a distinct and specific complex with its target RNA (rev-responsive element), a 234-nucleotide sequence within the envelope coding region of human immunodeficiency virus 1. No complex formation could be observed using RNAs with similar secondary structure nor with other human immunodeficiency virus 1 recombinant proteins. Deletion analysis mapped this specific binding to the first 90 nucleotides of this rev-responsive element, which contains a U2 small nuclear RNA homologous region. We propose that the specific binding of rev to its target RNA sequence plays an essential part in releasing an incompletely spliced viral mRNA containing this target sequence to the cytoplasm.
Daefler
S
S
Laboratory of Tumor Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Klotman
M E
ME
Wong-Staal
F
F
D001483:Base Sequence; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D015694:Gene Products, rev / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D012342:RNA, Small Nuclear / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D015534:Trans-Activators / Q000378:metabolism*; D015533:Transcriptional Activation; D014162:Transfection; D015678:Viral Structural Proteins / Q000235:genetics; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D015345:Oligonucleotide Probes; D012342:RNA, Small Nuclear; D012367:RNA, Viral; D011994:Recombinant Proteins; D015534:Trans-Activators; D015678:Viral Structural Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus