Hybrid Ty1/HIV-1 elements used to detect inhibitors and monitor the activity of HIV-1 reverse transcriptase

Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13905-10. doi: 10.1073/pnas.95.23.13905.


We previously demonstrated that hybrid retrotransposons composed of the yeast Ty1 element and the reverse transcriptase (RT) of HIV-1 are active in the yeast Saccharomyces cerevisiae. The RT activity of these hybrid Ty1/HIV-1 (his3AI/AIDS RT; HART) elements can be monitored by using a simple genetic assay. HART element reverse transcription depends on both the polymerase and RNase H domains of HIV-1 RT. Here we demonstrate that the HART assay is sensitive to inhibitors of HIV-1 RT. (-)-(S)-8-Chloro-4,5,6, 7-tetrahydro-5-methyl-6-(3-methyl-2-butenyl)imidazo[4,5,1-jk][1, 4]-benzodiazepin-2(1H)-thione monohydrochloride (8 Cl-TIBO), a well characterized non-nucleoside RT inhibitor (NNRTI) of HIV-1 RT, blocks propagation of HART elements. HART elements that express NNRTI-resistant RT variants of HIV-1 are insensitive to 8 Cl-TIBO, demonstrating the specificity of inhibition in this assay. HART elements carrying NNRTI-resistant variants of HIV-1 RT can be used to identify compounds that are active against drug-resistant viruses.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biological Assay
  • HIV Reverse Transcriptase / analysis*
  • HIV Reverse Transcriptase / genetics*
  • Humans
  • Recombinant Fusion Proteins / genetics
  • Retroelements / genetics*
  • Saccharomyces cerevisiae


  • Recombinant Fusion Proteins
  • Retroelements
  • HIV Reverse Transcriptase