Sequence specific cleavage of the HIV-1 coreceptor CCR5 gene by a hammer-head ribozyme and a DNA-enzyme: inhibition of the coreceptor function by DNA-enzyme

FEBS Lett. 1998 Oct 2;436(2):233-8. doi: 10.1016/s0014-5793(98)01137-5.


The chemokine receptor CCR5 is used as a major coreceptor for fusion and entry by non-syncytia inducing macrophage tropic isolates of HIV-1, which is mainly involved in transmission. Individuals who are homozygous for the delta32 allele of CCR5 are usually resistant to HIV-1 infection and continue to lead a normal healthy life. Thus this gene is dispensable and is, therefore, an attractive target in the host cell for interfering specifically with the virus-host interaction. With the aim to develop a specific antiviral approach at the molecular level, we have synthesized a hammer-head ribozyme and a DNA-enzyme. Both ribozyme and DNA-enzyme cleaved the CCR5 RNA in a sequence specific manner. This cleavage was protein independent but Mg2+ dependent. The extent of cleavage increased with increasing concentration of magnesium chloride. DNA-enzyme was more effective in cleaving a full length (1376 bases) in vitro generated transcript of CCR5 gene. In this communication, we show that the DNA-enzyme when introduced into a mammalian cell, results in decreased CD4-CCR5-gp160 mediated fusion of cell membranes. Potential applications of these trans acting molecules are discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • Cloning, Molecular
  • DNA / chemistry
  • DNA / metabolism*
  • Deoxyribonucleases, Type II Site-Specific / metabolism
  • HIV-1 / physiology
  • Humans
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Nucleic Acid Conformation
  • RNA, Catalytic / antagonists & inhibitors
  • RNA, Catalytic / chemistry*
  • RNA, Catalytic / metabolism*
  • Receptors, CCR5 / biosynthesis
  • Receptors, CCR5 / genetics*
  • Recombinant Proteins / biosynthesis
  • Substrate Specificity
  • Transcription, Genetic


  • RNA, Catalytic
  • Receptors, CCR5
  • Recombinant Proteins
  • DNA
  • endodeoxyribonuclease XBAI
  • Deoxyribonucleases, Type II Site-Specific