1. Academic Validation
  2. Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase

Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase

  • J Biol Chem. 2015 Jan 16;290(3):1474-84. doi: 10.1074/jbc.M114.614305.
Jean A Bernatchez 1 Rakesh Paul 2 Egor P Tchesnokov 3 Marianne Ngure 3 Greg L Beilhartz 3 Albert M Berghuis 4 Rico Lavoie 5 Lianhai Li 5 Anick Auger 5 Roman A Melnyk 5 Jay A Grobler 5 Michael D Miller 5 Daria J Hazuda 5 Sidney M Hecht 2 Matthias Götte 6
Affiliations

Affiliations

  • 1 From the Department of Biochemistry, McGill University, Montreal, Quebec H3G 1Y6, Canada.
  • 2 the Biodesign Institute and Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287.
  • 3 the Department of Microbiology and Immunology, McGill University, Montreal, Quebec H3A 2B4, Canada.
  • 4 From the Department of Biochemistry, McGill University, Montreal, Quebec H3G 1Y6, Canada, the Department of Microbiology and Immunology, McGill University, Montreal, Quebec H3A 2B4, Canada.
  • 5 Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
  • 6 From the Department of Biochemistry, McGill University, Montreal, Quebec H3G 1Y6, Canada, the Department of Microbiology and Immunology, McGill University, Montreal, Quebec H3A 2B4, Canada, the Department of Medicine, Division of Experimental Medicine, McGill University, Quebec H3A 1A3, Canada gotte@ualberta.ca.
Abstract

The pyrophosphate mimic and broad spectrum Antiviral phosphonoformic acid (PFA, foscarnet) was shown to freeze the pre-translocational state of the Reverse Transcriptase (RT) complex of the human immunodeficiency virus type 1 (HIV-1). However, PFA lacks a specificity domain, which is seen as a major reason for toxic side effects associated with the clinical use of this drug. Here, we studied the mechanism of inhibition of HIV-1 RT by the 4-chlorophenylhydrazone of mesoxalic acid (CPHM) and demonstrate that this compound also blocks RT translocation. Hot spots for inhibition with PFA or CPHM occur at template positions with a bias toward pre-translocation. Mutations at active site residue Asp-185 compromise binding of both compounds. Moreover, divalent metal ions are required for the formation of ternary complexes with either of the two compounds. However, CPHM contains both an anchor domain that likely interacts with the catalytic metal ions and a specificity domain. Thus, although the inhibitor binding sites may partly overlap, they are not identical. The K65R mutation in HIV-1 RT, which reduces affinity to PFA, increases affinity to CPHM. Details with respect to the binding sites of the two inhibitors are provided on the basis of mutagenesis studies, structure-activity relationship analyses with newly designed CPHM derivatives, and in silico docking experiments. Together, these findings validate the pre-translocated complex of HIV-1 RT as a specific target for the development of novel classes of RT inhibitors.

Keywords

Antiviral Agent; Drug Action; Drug Resistance; Human Immunodeficiency Virus (HIV); Reverse Transcription.

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