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

Jean A. Bernatchez, Rakesh Paul, Egor P. Tchesnokov, Marianne Ngure, Greg L. Beilhartz, Albert M. Berghuis, Rico Lavoie, Lianhai Li, Anick Auger, Roman A. Melnyk, Jay A. Grobler, Michael D. Miller, Daria J. Hazuda, Sidney Hecht, Matthias Götte

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1474-1484
Number of pages11
JournalJournal of Biological Chemistry
Volume290
Issue number3
DOIs
StatePublished - Jan 16 2015

Fingerprint

HIV-1
Foscarnet
Derivatives
RNA-Directed DNA Polymerase
Metal ions
Metals
Binding Sites
Ions
Mutagenesis
Mutation
Reverse Transcriptase Inhibitors
Poisons
Structure-Activity Relationship
Anchors
Viruses
Computer Simulation
Antiviral Agents
Catalytic Domain
mesoxalic acid
Human immunodeficiency virus 1 reverse transcriptase

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Bernatchez, J. A., Paul, R., Tchesnokov, E. P., Ngure, M., Beilhartz, G. L., Berghuis, A. M., ... Götte, M. (2015). Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase. Journal of Biological Chemistry, 290(3), 1474-1484. https://doi.org/10.1074/jbc.M114.614305

Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase. / Bernatchez, Jean A.; Paul, Rakesh; Tchesnokov, Egor P.; Ngure, Marianne; Beilhartz, Greg L.; Berghuis, Albert M.; Lavoie, Rico; Li, Lianhai; Auger, Anick; Melnyk, Roman A.; Grobler, Jay A.; Miller, Michael D.; Hazuda, Daria J.; Hecht, Sidney; Götte, Matthias.

In: Journal of Biological Chemistry, Vol. 290, No. 3, 16.01.2015, p. 1474-1484.

Research output: Contribution to journalArticle

Bernatchez, JA, Paul, R, Tchesnokov, EP, Ngure, M, Beilhartz, GL, Berghuis, AM, Lavoie, R, Li, L, Auger, A, Melnyk, RA, Grobler, JA, Miller, MD, Hazuda, DJ, Hecht, S & Götte, M 2015, 'Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase', Journal of Biological Chemistry, vol. 290, no. 3, pp. 1474-1484. https://doi.org/10.1074/jbc.M114.614305
Bernatchez JA, Paul R, Tchesnokov EP, Ngure M, Beilhartz GL, Berghuis AM et al. Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase. Journal of Biological Chemistry. 2015 Jan 16;290(3):1474-1484. https://doi.org/10.1074/jbc.M114.614305
Bernatchez, Jean A. ; Paul, Rakesh ; Tchesnokov, Egor P. ; Ngure, Marianne ; Beilhartz, Greg L. ; Berghuis, Albert M. ; Lavoie, Rico ; Li, Lianhai ; Auger, Anick ; Melnyk, Roman A. ; Grobler, Jay A. ; Miller, Michael D. ; Hazuda, Daria J. ; Hecht, Sidney ; Götte, Matthias. / Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase. In: Journal of Biological Chemistry. 2015 ; Vol. 290, No. 3. pp. 1474-1484.
@article{aa22be95b4234b81a7b95473090a3977,
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.",
author = "Bernatchez, {Jean A.} and Rakesh Paul and Tchesnokov, {Egor P.} and Marianne Ngure and Beilhartz, {Greg L.} and Berghuis, {Albert M.} and Rico Lavoie and Lianhai Li and Anick Auger and Melnyk, {Roman A.} and Grobler, {Jay A.} and Miller, {Michael D.} and Hazuda, {Daria J.} and Sidney Hecht and Matthias G{\"o}tte",
year = "2015",
month = "1",
day = "16",
doi = "10.1074/jbc.M114.614305",
language = "English (US)",
volume = "290",
pages = "1474--1484",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "3",

}

TY - JOUR

T1 - Derivatives of mesoxalic acid block translocation of HIV-1 reverse transcriptase

AU - Bernatchez, Jean A.

AU - Paul, Rakesh

AU - Tchesnokov, Egor P.

AU - Ngure, Marianne

AU - Beilhartz, Greg L.

AU - Berghuis, Albert M.

AU - Lavoie, Rico

AU - Li, Lianhai

AU - Auger, Anick

AU - Melnyk, Roman A.

AU - Grobler, Jay A.

AU - Miller, Michael D.

AU - Hazuda, Daria J.

AU - Hecht, Sidney

AU - Götte, Matthias

PY - 2015/1/16

Y1 - 2015/1/16

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84922228750&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84922228750&partnerID=8YFLogxK

U2 - 10.1074/jbc.M114.614305

DO - 10.1074/jbc.M114.614305

M3 - Article

VL - 290

SP - 1474

EP - 1484

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -