No tRNA3Lys unwinding in a complex with HIV NCp7

J Biol Chem. 1997 Oct 3;272(40):25143-8. doi: 10.1074/jbc.272.40.25143.

Abstract

The nucleocapsid protein NCp7 of the human immunodeficiency virus (HIV) type 1 is important for the annealing of HIV RNA and tRNA3Lys, the tRNA acting as a primer during reverse transcription of HIV RNA. A wild type NCp7 and a Cys23 mutant having a disrupted zinc finger were analyzed with far UV circular dichroism (CD). CD data analysis revealed that NCp7 has a high content of extended structures in aqueous buffer, decreasing in Cys23 NCp7 and in NCp7 in the absence of zinc. An increase in beta-turn structures is observed in NCp7 bound to tRNA3Lys. Furthermore, CD data shows that Cys23 NCp7 binds tRNA3Lys. The CD spectrum of tRNA3Lys is typical of an A-form helix and retains this structure after binding of NCp7, which demonstrates that NCp7 does not induce tRNA3Lys unwinding. CD spectra of tRNA3Lys were measured from 5 to 80 degrees C to observe CD changes resulting from tRNA3Lys melting. Molecular modeling of the complex identifies two potential tRNA anticodon binding sites in the NCp7 N-terminal region and first zinc finger. In this model, both binding sites can interact with 12 nucleotides in the anticodon domain without requiring a base specificity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Capsid / chemistry*
  • Capsid / metabolism*
  • Capsid Proteins*
  • Circular Dichroism
  • Gene Products, gag / chemistry*
  • Gene Products, gag / metabolism*
  • HIV-1 / genetics
  • HIV-1 / metabolism*
  • Humans
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleic Acid Conformation*
  • Protein Structure, Secondary*
  • Spectrophotometry, Ultraviolet
  • Viral Proteins*
  • Zinc Fingers
  • gag Gene Products, Human Immunodeficiency Virus

Substances

  • Capsid Proteins
  • Gene Products, gag
  • NCP7 protein, Human immunodeficiency virus 1
  • Viral Proteins
  • gag Gene Products, Human Immunodeficiency Virus