A rev/beta-galactosidase fusion protein binds in vitro transcripts spanning the rev-responsive element of human immunodeficiency virus type 1 (HIV-1).
The rev protein of human immunodeficiency virus type 1 (HIV-1), a phosphoprotein of 20 kDa apparent molecular mass, is essential to target the mRNA for virion polypeptides into the cytoplasm. This effect is mediated by a specific RNA stretch (rev-responsive element = RRE) localized within a 3'-terminal segment of the mRNA encoding virion proteins. We present evidence that rev expressed as a beta-galactosidase fusion protein in E. coli forms a complex with in vitro transcripts containing the RRE; it can be precipitated by monoclonal antibodies with rev or beta-galactosidase specificity. In addition, specific binding of rev protein to RNA could be demonstrated by Northwestern blotting.
Aepinus
C
C
Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, FRG.
Voll
R
R
Bröker
M
M
Fleckenstein
B
B
D000818:Animals; D000911:Antibodies, Monoclonal; D005696:Galactosidases / Q000378:metabolism*; D015694:Gene Products, rev / Q000235:genetics / Q000378:metabolism*; D005814:Genes, Viral; D015757:Genes, rev; D015497:HIV-1 / Q000235:genetics*; D015151:Immunoblotting; D051379:Mice; D011233:Precipitin Tests; D012367:RNA, Viral / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D015534:Trans-Activators / Q000378:metabolism*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000378:metabolism*; D054321:rev Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; D015694:Gene Products, rev; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D054321:rev Gene Products, Human Immunodeficiency Virus; D005696:Galactosidases; D001616:beta-Galactosidase