The N137 and P140 amino acids in the p51 and the P95 amino acid in the p66 subunit of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase are instrumental to maintain catalytic activity and to design new classes of anti-HIV-1 drugs

Joeri Auwerx, Joke Van Nieuwenhove, Fátima Rodríguez-Barrios, Sonia De Castro, Sonsoles Velázquez, Francesca Ceccherini-Silberstein, Erik De Clercq, María José Camarasa, Carlo Federico Perno, Federico Gago, Jan Balzarini

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Amino acids N137 and P140 in the p51 subunit of HIV-1 reverse transcriptase (RT) are part of the β7-β8-loop that contributes to the formation of the base of the non-nucleoside RT inhibitor (NNRTI)-binding pocket and makes up a substantial part of the dimerization interface. Amino acid P95 in p66 also markedly contributes to the dimerization binding energy. Nine RT mutants at amino acid 137 were constructed bearing the mutations Y, K, T, D, A, Q, S, H or E. The prolines at amino acid positions 95 and 140 were replaced by alanine in separate enzymes. We found that all mutant RT enzymes showed a dramatically decreased RNA-dependent DNA polymerase activity. None of the mutant RT enzymes showed marked resistance against any of the clinically used NNRTIs but they surprisingly lost significant sensitivity for NRTIs such as ddGTP. The denaturation analyses of the mutant RTs by urea are suggestive for a relevant role of N137 in the stability of the RT heterodimer and support the view that the β7-β8 loop in p51 is a hot spot for RT dimerization and instrumental for efficient polymerase catalytic activity. Consequently, N137 and P140 in p51 and P95 in p66 should be attractive targets in the design of new structural classes of RT inhibitors aimed at compromising the optimal interaction of the β7-β8 loop in p51 at the p66/p51 dimerization interface.

Original languageEnglish
Pages (from-to)2294-2300
Number of pages7
JournalFEBS Letters
Volume579
Issue number11
DOIs
Publication statusPublished - Apr 25 2005

Fingerprint

RNA-Directed DNA Polymerase
Viruses
HIV-1
Catalyst activity
Dimerization
Amino Acids
Pharmaceutical Preparations
Reverse Transcriptase Inhibitors
Bearings (structural)
Enzymes
Denaturation
Binding energy
Proline
Alanine
Urea
Human immunodeficiency virus 1 reverse transcriptase
Mutation

Keywords

  • Anti-HIV-1 drugs
  • HIV-1
  • p51
  • p66

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

The N137 and P140 amino acids in the p51 and the P95 amino acid in the p66 subunit of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase are instrumental to maintain catalytic activity and to design new classes of anti-HIV-1 drugs. / Auwerx, Joeri; Van Nieuwenhove, Joke; Rodríguez-Barrios, Fátima; De Castro, Sonia; Velázquez, Sonsoles; Ceccherini-Silberstein, Francesca; De Clercq, Erik; Camarasa, María José; Perno, Carlo Federico; Gago, Federico; Balzarini, Jan.

In: FEBS Letters, Vol. 579, No. 11, 25.04.2005, p. 2294-2300.

Research output: Contribution to journalArticle

Auwerx, Joeri ; Van Nieuwenhove, Joke ; Rodríguez-Barrios, Fátima ; De Castro, Sonia ; Velázquez, Sonsoles ; Ceccherini-Silberstein, Francesca ; De Clercq, Erik ; Camarasa, María José ; Perno, Carlo Federico ; Gago, Federico ; Balzarini, Jan. / The N137 and P140 amino acids in the p51 and the P95 amino acid in the p66 subunit of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase are instrumental to maintain catalytic activity and to design new classes of anti-HIV-1 drugs. In: FEBS Letters. 2005 ; Vol. 579, No. 11. pp. 2294-2300.
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AU - Auwerx, Joeri

AU - Van Nieuwenhove, Joke

AU - Rodríguez-Barrios, Fátima

AU - De Castro, Sonia

AU - Velázquez, Sonsoles

AU - Ceccherini-Silberstein, Francesca

AU - De Clercq, Erik

AU - Camarasa, María José

AU - Perno, Carlo Federico

AU - Gago, Federico

AU - Balzarini, Jan

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