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Hybrid Ty1/Hiv-1 Elements Used to Detect Inhibitors and Monitor the Activity of Hiv-1 Reverse Transcriptase

  1. Author:
    Nissley, D. V.
    Boyer, P. L.
    Garfinkel, D. J.
    Hughes, S. H.
    Strathern, J. N.

  2. Author Address

    Nissley DV NCI GENE REGULAT & CHROMOSOME BIOL LAB FREDERICK CANC RES & DEV CTR ABL BASIC RES PROGRAM FREDERICK, MD 21702 USA NCI MOL BIOL CARCINOGENESIS LAB FREDERICK CANC RES & DEV CTR ABL BASIC RES PROGRAM FREDERICK, MD 21702 USA
    1. Year: 1998
  2. Journal: Proceedings of the National Academy of Sciences of the United States of America
    1. 95
    2. 23
    3. Pages: 13905-13910
  4. Type of Article: Article
  1. Abstract:

    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 (his3A1/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)imid azo [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. [References: 58]

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