title: Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase.
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.
authors.affiliation: From the Department of Biochemistry, McGill University, Montreal, Quebec H3G 1Y6, Canada.
authors.forename: Jean A
authors.initials: JA
authors.lastname: Bernatchez
authors.affiliation: the Biodesign Institute and Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287.
authors.forename: Rakesh
authors.initials: R
authors.lastname: Paul
authors.affiliation: the Department of Microbiology and Immunology, McGill University, Montreal, Quebec H3A 2B4, Canada.
authors.forename: Egor P
authors.initials: EP
authors.lastname: Tchesnokov
authors.affiliation: the Department of Microbiology and Immunology, McGill University, Montreal, Quebec H3A 2B4, Canada.
authors.forename: Marianne
authors.initials: M
authors.lastname: Ngure
authors.affiliation: the Department of Microbiology and Immunology, McGill University, Montreal, Quebec H3A 2B4, Canada.
authors.forename: Greg L
authors.initials: GL
authors.lastname: Beilhartz
authors.affiliation: 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.
authors.forename: Albert M
authors.initials: AM
authors.lastname: Berghuis
authors.affiliation: Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
authors.forename: Rico
authors.initials: R
authors.lastname: Lavoie
authors.affiliation: Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
authors.forename: Lianhai
authors.initials: L
authors.lastname: Li
authors.affiliation: Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
authors.forename: Anick
authors.initials: A
authors.lastname: Auger
authors.affiliation: Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
authors.forename: Roman A
authors.initials: RA
authors.lastname: Melnyk
authors.affiliation: Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
authors.forename: Jay A
authors.initials: JA
authors.lastname: Grobler
authors.affiliation: Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Miller
authors.affiliation: Merck Research Laboratories, West Point, Pennsylvania 19486-8000, and.
authors.forename: Daria J
authors.initials: DJ
authors.lastname: Hazuda
authors.affiliation: the Biodesign Institute and Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287.
authors.forename: Sidney M
authors.initials: SM
authors.lastname: Hecht
authors.affiliation: 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.
authors.forename: Matthias
authors.initials: M
authors.lastname: Götte
mesh_terms: D044966:Anti-Retroviral Agents / Q000737:chemistry; D002384:Catalysis; D020134:Catalytic Domain; D004353:Drug Evaluation, Preclinical; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors*; D015497:HIV-1 / Q000201:enzymology*; D006835:Hydrazones / Q000737:chemistry*; D007477:Ions; D008314:Malonates / Q000737:chemistry*; D008670:Metals / Q000737:chemistry; D008958:Models, Molecular; D016296:Mutagenesis; D009154:Mutation; D011485:Protein Binding; D055503:Protein Multimerization; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry*; D016914:Ribonuclease H / Q000737:chemistry; D013329:Structure-Activity Relationship
chemical_list: C417774:4-chlorophenylhydrazone mesoxalic acid; D044966:Anti-Retroviral Agents; D006835:Hydrazones; D007477:Ions; D008314:Malonates; D008670:Metals; D018894:Reverse Transcriptase Inhibitors; D054303:HIV Reverse Transcriptase; D016914:Ribonuclease H
keywords: Antiviral Agent; Drug Action; Drug Resistance; Human Immunodeficiency Virus (HIV); Reverse Transcription