A rev/beta-galactosidase fusion protein binds in vitro transcripts spanning the rev-responsive element of human immunodeficiency virus type 1 (HIV-1)

FEBS Lett. 1990 Apr 24;263(2):217-21. doi: 10.1016/0014-5793(90)81377-z.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Galactosidases / metabolism*
  • Gene Products, rev / genetics
  • Gene Products, rev / metabolism*
  • Genes, Viral*
  • Genes, rev*
  • HIV-1 / genetics*
  • Immunoblotting
  • Mice
  • Precipitin Tests
  • RNA, Viral / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Regulatory Sequences, Nucleic Acid
  • Trans-Activators / metabolism*
  • Transcription, Genetic
  • beta-Galactosidase / metabolism*
  • rev Gene Products, Human Immunodeficiency Virus

Substances

  • Antibodies, Monoclonal
  • Gene Products, rev
  • RNA, Viral
  • Recombinant Fusion Proteins
  • Trans-Activators
  • rev Gene Products, Human Immunodeficiency Virus
  • Galactosidases
  • beta-Galactosidase